MAD2L1

id: Q13257
gene_symbol: MAD2L1
aliases:
- MAD2
- HSMAD2
- REV7
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: >-
  MAD2L1 (Mitotic Arrest Deficient 2-Like 1) encodes a HORMA domain-containing protein
  that is a core effector of the spindle assembly checkpoint (SAC). MAD2 exists in
  two
  interconvertible conformations: an open form (O-MAD2) that predominates in the cytosol
  and a closed form (C-MAD2) that binds its key targets. At unattached kinetochores
  during
  prometaphase, MAD1:C-MAD2 core complexes recruit cytosolic O-MAD2 and catalyze its
  conversion to C-MAD2, enabling binding to CDC20. The resulting CDC20:MAD2 complex
  associates with BUB3 and BUBR1 to form the mitotic checkpoint complex (MCC), which
  inhibits the anaphase-promoting complex/cyclosome (APC/C-CDC20), thereby preventing
  premature anaphase until all chromosomes achieve proper bipolar attachment. Checkpoint
  silencing is mediated by p31comet (MAD2L1BP) and the AAA+ ATPase TRIP13, which
  remodel C-MAD2 back to O-MAD2 and disassemble the MCC. MAD2 also localizes to the
  nuclear pore complex via TPR during interphase, potentially facilitating early MCC
  assembly.

existing_annotations:
- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  review:
    summary: >-
      MAD2 localization to kinetochores is well-established and central to its checkpoint
      function. The original discovery paper (PMID:8824189) showed MAD2 localizes
      to
      kinetochores after chromosome condensation but is absent from kinetochores
      at
      metaphase.
    action: ACCEPT
    reason: >-
      Kinetochore localization is a core aspect of MAD2 function. IBA inference
      from
      phylogenetic analysis is consistent with abundant experimental evidence showing
      MAD2 recruitment to unattached kinetochores via the MAD1:MAD2 core complex.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "Human, or Homo sapiens, MAD2 (hsMAD2) was localized at the
        kinetochore after chromosome condensation but was no longer observed at the
        kinetochore in metaphase, suggesting that MAD2 might monitor the completeness
        of the spindle-kinetochore attachment"
    - reference_id: PMID:18981471
      supporting_text: "Depletion of Tpr decreases the levels of Mad1 at kinetochores
        during prometaphase, correlating with the inability of Mad1 to activate Mad2"

    - reference_id: file:human/MAD2L1/MAD2L1-deep-research-falcon.md
      supporting_text: 'model: Edison Scientific Literature'
- term:
    id: GO:0007094
    label: mitotic spindle assembly checkpoint signaling
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  review:
    summary: >-
      MAD2 is a core component of the spindle assembly checkpoint. The deep research
      confirms
      that MAD2 is "a core effector of the spindle assembly checkpoint (SAC)" and
      is
      essential for MCC formation and APC/C inhibition.
    action: ACCEPT
    reason: >-
      This is the primary biological process for MAD2. The IBA annotation is fully
      consistent
      with decades of research establishing MAD2 as essential for SAC signaling.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "The human homolog of MAD2 was isolated and shown to be a necessary
        component of the mitotic checkpoint in HeLa cells by antibody electroporation
        experiments"
    - reference_id: PMID:18981471
      supporting_text: "The mitotic arrest-deficient protein Mad1 forms a complex
        with Mad2, which is required for imposing mitotic arrest on cells in which
        the spindle assembly is perturbed"

- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  review:
    summary: >-
      Computational annotation to kinetochore based on combined automated methods.
      Duplicates
      the IBA annotation above.
    action: ACCEPT
    reason: >-
      Consistent with the IBA annotation and extensive experimental evidence. Redundant
      annotations with different evidence types are acceptable.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "Human, or Homo sapiens, MAD2 (hsMAD2) was localized at the
        kinetochore after chromosome condensation but was no longer observed at the
        kinetochore in metaphase"

- term:
    id: GO:0000922
    label: spindle pole
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  review:
    summary: >-
      UniProt subcellular location annotation indicates spindle pole localization.
      The deep
      research mentions MAD2 presence at spindle poles as part of mitotic checkpoint
      dynamics.
    action: ACCEPT
    reason: >-
      Spindle pole localization is documented in UniProt and consistent with MAD2's
      role in
      mitotic checkpoint function. This represents a secondary localization site
      during mitosis.
    supported_by:
    - reference_id: PMID:20133940
      supporting_text: "Here, we show association of another nucleoporin, termed Tpr
        (translocated promoter region), with the molecular motors dynein and dynactin,
        which both orchestrate with the spindle checkpoints Mad1 and Mad2 during cell
        division"

- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  review:
    summary: >-
      Nuclear localization is inferred from automated methods. UniProt confirms
      nuclear
      localization through IDA evidence.
    action: ACCEPT
    reason: >-
      Nuclear localization is experimentally validated (see IDA annotations below).
      MAD2
      is recruited to the nuclear pore complex by TPR during interphase.
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "Tpr directly binds to Mad1 and Mad2...Depletion of Tpr in
        HeLa cells disrupts the NPC localization of Mad1 and Mad2 during interphase"

- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  review:
    summary: >-
      Cytoplasmic localization is inferred from automated methods. The open form
      of MAD2
      (O-MAD2) exists as a cytosolic pool that is recruited to kinetochores.
    action: ACCEPT
    reason: >-
      The cytosolic O-MAD2 pool is essential for the template model of MAD2 activation.
      Cytoplasmic localization is well-documented.
    supported_by:
    - reference_id: PMID:19010891
      supporting_text: "MAD1beta localized in the cytoplasm...MAD1beta was found to
        physically interact with MAD2 and sequester it in the cytoplasm"

- term:
    id: GO:0051301
    label: cell division
  evidence_type: IEA
  original_reference_id: GO_REF:0000043
  review:
    summary: >-
      Inferred from UniProt keyword "Cell division". MAD2 function in the spindle
      checkpoint
      is essential for proper cell division.
    action: ACCEPT
    reason: >-
      This is a broader process that encompasses the SAC. MAD2's role in preventing
      premature anaphase is essential for accurate chromosome segregation during
      cell division.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "In Saccharomyces cerevisiae, MAD2 is required for mitotic
        arrest if the spindle assembly is perturbed"

- term:
    id: GO:0051783
    label: regulation of nuclear division
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  review:
    summary: >-
      ARBA machine learning annotation for regulation of nuclear division. MAD2
      regulates
      nuclear division by controlling the metaphase-to-anaphase transition.
    action: ACCEPT
    reason: >-
      This is an appropriate parent term for MAD2's role in checkpoint signaling.
      MAD2-mediated MCC formation regulates the timing of anaphase onset.
    supported_by:
    - reference_id: PMID:10700282
      supporting_text: "The checkpoint protein Mad2 inhibits the activity of the anaphase
        promoting complex by sequestering Cdc20 until all chromosomes are aligned
        at the metaphase plate"

- term:
    id: GO:1901991
    label: negative regulation of mitotic cell cycle phase transition
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  review:
    summary: >-
      ARBA annotation for negative regulation of mitotic cell cycle phase transition.
      MAD2 inhibits the metaphase-to-anaphase transition via APC/C inhibition.
    action: ACCEPT
    reason: >-
      Accurate annotation. MAD2 negatively regulates the metaphase-to-anaphase transition
      by inhibiting APC/C-CDC20 activity through MCC formation.
    supported_by:
    - reference_id: PMID:10700282
      supporting_text: "The checkpoint protein Mad2 inhibits the activity of the anaphase
        promoting complex by sequestering Cdc20"

- term:
    id: GO:1990728
    label: mitotic spindle assembly checkpoint MAD1-MAD2 complex
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  review:
    summary: >-
      ARBA annotation for membership in the MAD1-MAD2 complex. This is a core structural
      complex essential for MAD2 activation and checkpoint signaling.
    action: ACCEPT
    reason: >-
      Highly accurate annotation. The MAD1:C-MAD2 core complex at kinetochores is
      essential
      for templated conversion of O-MAD2 to C-MAD2 and subsequent CDC20 capture.
    supported_by:
    - reference_id: PMID:12006501
      supporting_text: "The crystal structure of the Mad1-Mad2 complex reveals an
        asymmetric tetramer, with elongated Mad1 monomers parting from a coiled-coil
        to form two connected sub-complexes with Mad2"
    - reference_id: PMID:18981471
      supporting_text: "The mitotic arrest-deficient protein Mad1 forms a complex
        with Mad2"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:10527948
  review:
    summary: >-
      Interaction with ADAM17/TACE demonstrated by immunoprecipitation. This is
      an unusual
      interaction partner for a checkpoint protein.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      While the interaction is documented, "protein binding" is too vague. The interaction
      with ADAM17 is not clearly related to MAD2's core checkpoint function and
      may represent
      a peripheral or indirect interaction.
    supported_by:
    - reference_id: PMID:10527948
      supporting_text: "Evidence for an interaction of the metalloprotease-disintegrin
        tumour necrosis factor alpha convertase (TACE) with mitotic arrest deficient
        2 (MAD2)"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:15182668
  review:
    summary: >-
      Interaction with CDC20 (Q12834) documented. This is a core functional interaction
      for checkpoint signaling.
    action: MODIFY
    reason: >-
      The interaction with CDC20 is the primary functional output of MAD2 activation.
      "Protein binding" is too vague; this should be annotated to the specific complex
      or with a more informative term.
    proposed_replacement_terms:
    - id: GO:1990333
      label: mitotic checkpoint complex, CDC20-MAD2 subcomplex
    supported_by:
    - reference_id: PMID:10700282
      supporting_text: "Mad2 possesses a novel three-layered alpha/beta fold with
        three alpha-helices packed between two beta-sheets...Mad2 and Cdc20 form a
        tight 1:1 heterodimeric complex"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:16189514
  review:
    summary: >-
      High-throughput protein interaction study. Interactions with TSC22D4 and MAD1L1
      documented. MAD1L1 interaction is core; TSC22D4 significance unclear.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Generic protein binding annotation from HTP study. The MAD1L1 interaction
      is captured
      in more specific annotations.
    supported_by:
    - reference_id: PMID:16189514
      supporting_text: "Towards a proteome-scale map of the human protein-protein
        interaction network"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:16525508
  review:
    summary: >-
      Interaction with CDC20 documented in study of MAD2 conformational dimerization.
    action: MODIFY
    reason: >-
      This study specifically addresses MAD2-CDC20 interaction in the context of
      conformational dimerization and p31comet inhibition. Should be annotated to
      more
      specific complex terms.
    proposed_replacement_terms:
    - id: GO:1990333
      label: mitotic checkpoint complex, CDC20-MAD2 subcomplex
    supported_by:
    - reference_id: PMID:16525508
      supporting_text: "Determinants of conformational dimerization of Mad2 and its
        inhibition by p31comet"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:17443180
  review:
    summary: >-
      Interactions with CDC27 (P30260) and CDC20 (Q12834) documented in study of
      ubiquitination/deubiquitination in anaphase initiation.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Generic protein binding. CDC27 is an APC/C subunit; interaction likely reflects
      MCC-APC/C association. Core CDC20 interaction captured elsewhere.
    supported_by:
    - reference_id: PMID:17443180
      supporting_text: "Anaphase initiation is regulated by antagonistic ubiquitination
        and deubiquitination activities"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:17443186
  review:
    summary: >-
      Interactions with CDC27 and CDC20 documented in study of spindle checkpoint
      inactivation.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Duplicate of interactions captured in more specific annotations. Generic protein
      binding is not informative.
    supported_by:
    - reference_id: PMID:17443186
      supporting_text: "Ubiquitination by the anaphase-promoting complex drives spindle
        checkpoint inactivation"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:18022367
  review:
    summary: >-
      Interaction with CDC20 demonstrated in seminal structural study of the MAD2
      conformational dimer.
    action: MODIFY
    reason: >-
      This landmark paper establishes the structural basis for O-MAD2/C-MAD2 dimerization
      and CDC20 binding. Should be annotated to specific complex terms.
    proposed_replacement_terms:
    - id: GO:1990333
      label: mitotic checkpoint complex, CDC20-MAD2 subcomplex
    supported_by:
    - reference_id: PMID:18022367
      supporting_text: "The structure of the O-Mad2-C-Mad2 conformational dimer is
        consistent with a catalytic model in which a C-Mad2 template facilitates the
        binding of O-Mad2 to Cdc20"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:18022368
  review:
    summary: >-
      Interaction with p31comet (MAD2L1BP, Q15013) demonstrated. p31comet is the
      key
      negative regulator of MAD2 that promotes checkpoint silencing.
    action: MODIFY
    reason: >-
      This is a functionally important interaction for checkpoint silencing. p31comet
      blocks MAD2 activation through structural mimicry. Should be annotated to
      a more
      specific MF term if available.
    proposed_replacement_terms:
    - id: GO:0042802
      label: identical protein binding
    supported_by:
    - reference_id: PMID:18022368
      supporting_text: "p31comet blocks Mad2 activation through structural mimicry"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:18318601
  review:
    summary: >-
      Interactions with CDC20 and MAD1L1 demonstrated in structural study of symmetric
      MAD2 dimer.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured in more specific annotations. This study addresses
      homodimerization which is separately annotated.
    supported_by:
    - reference_id: PMID:18318601
      supporting_text: "Insights into mad2 regulation in the spindle checkpoint revealed
        by the crystal structure of the symmetric mad2 dimer"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:18692475
  review:
    summary: >-
      Interaction with MAD1L1 from C. elegans embryogenesis domain-based interactome.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Generic protein binding from interactome study. MAD1-MAD2 interaction is well
      captured in more specific annotations.
    supported_by:
    - reference_id: PMID:18692475
      supporting_text: "A protein domain-based interactome network for C. elegans
        early embryogenesis"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19143472
  review:
    summary: >-
      Interaction with CDC20 in study of Mad2 activation dynamics.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core CDC20 interaction captured elsewhere. Generic protein binding is not
      informative.
    supported_by:
    - reference_id: PMID:19143472
      supporting_text: "The influence of catalysis on mad2 activation dynamics"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19615732
  review:
    summary: >-
      Interaction with KEAP1 from deubiquitinating enzyme interactome study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Interaction with KEAP1 is not clearly related to core checkpoint function.
      May represent an indirect or peripheral interaction.
    supported_by:
    - reference_id: PMID:19615732
      supporting_text: "Defining the human deubiquitinating enzyme interaction landscape"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:20212161
  review:
    summary: >-
      Interactions with BUB1B, CDC27, and CDC20 documented in study of ATP-dependent
      APC/C release from MCC inhibition.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      These interactions are core to MCC function but captured in more specific
      annotations.
      BUB1B (BUBR1) is an MCC component.
    supported_by:
    - reference_id: PMID:20212161
      supporting_text: "ATP is required for the release of the anaphase-promoting
        complex/cyclosome from inhibition by the mitotic checkpoint"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:20360068
  review:
    summary: >-
      Interaction with CDC20 from systematic analysis of chromosome segregation
      proteins.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core CDC20 interaction captured elsewhere. HTP study.
    supported_by:
    - reference_id: PMID:20360068
      supporting_text: "Systematic analysis of human protein complexes identifies
        chromosome segregation proteins"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:20951947
  review:
    summary: >-
      Interaction with CDC27 from study of APC/C pharmacologic inhibition.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      CDC27 interaction reflects MCC-APC/C association. Captured in pathway annotations.
    supported_by:
    - reference_id: PMID:20951947
      supporting_text: "Pharmacologic inhibition of the anaphase-promoting complex
        induces a spindle checkpoint-dependent mitotic arrest"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21041666
  review:
    summary: >-
      Interactions with CDC20 and MAD1L1 documented in study of MAD2 phosphorylation
      and conformational transition.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere. Study addresses phosphoregulation of
      MAD2.
    supported_by:
    - reference_id: PMID:21041666
      supporting_text: "Phosphorylation of the spindle checkpoint protein Mad2 regulates
        its conformational transition"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21300909
  review:
    summary: >-
      Interactions with BUB1B, CDC27, and CDC20 in study of p31comet-mediated MCC
      disassembly.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core MCC interactions captured elsewhere.
    supported_by:
    - reference_id: PMID:21300909
      supporting_text: "p31comet Promotes disassembly of the mitotic checkpoint complex
        in an ATP-dependent process"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21407176
  review:
    summary: >-
      Interaction with CDC20 in study of Aurora B role in checkpoint signaling.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core CDC20 interaction captured elsewhere.
    supported_by:
    - reference_id: PMID:21407176
      supporting_text: "Evidence that Aurora B is implicated in spindle checkpoint
        signalling independently of error correction"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21666598
  review:
    summary: >-
      Interactions with p31comet and SGO2 documented. SGO2 as a MAD1/CDC20-like
      interactor.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      p31comet interaction captured elsewhere. SGO2 interaction may represent a
      regulatory
      mechanism but is not core to checkpoint function.
    supported_by:
    - reference_id: PMID:21666598
      supporting_text: "Shugoshin is a Mad1/Cdc20-like interactor of Mad2"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21772247
  review:
    summary: >-
      Multiple interactions (BUB1B, CDC27, CDC20, p31comet, MAD1L1) from study probing
      Mad1:C-Mad2 function in vivo.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured in specific annotations. HTP validation study.
    supported_by:
    - reference_id: PMID:21772247
      supporting_text: "Probing the in vivo function of Mad1:C-Mad2 in the spindle
        assembly checkpoint"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21988832
  review:
    summary: >-
      Interaction with MAD1L1 from liver protein interactome study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations. HTP study.
    supported_by:
    - reference_id: PMID:21988832
      supporting_text: "Toward an understanding of the protein interaction network
        of the human liver"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:22000412
  review:
    summary: >-
      Interaction with BUB1B from structural study of Blinkin-BUBR1 complex.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      BUB1B/BUBR1 is an MCC component. Interaction captured in MCC annotations.
    supported_by:
    - reference_id: PMID:22000412
      supporting_text: "Structure of a Blinkin-BUBR1 complex reveals an interaction
        crucial for kinetochore-mitotic checkpoint regulation"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:22340593
  review:
    summary: >-
      Interaction with CDC20 from study of Aurora kinase-A and p73.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core CDC20 interaction captured elsewhere.
    supported_by:
    - reference_id: PMID:22340593
      supporting_text: "Aurora kinase-A inactivates DNA damage-induced apoptosis and
        spindle assembly checkpoint response functions of p73"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:22493223
  review:
    summary: >-
      Interaction with MAD1L1 from structural study of MAD1 C-terminal domain.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations.
    supported_by:
    - reference_id: PMID:22493223
      supporting_text: "Structure of human Mad1 C-terminal domain reveals its involvement
        in kinetochore targeting"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:24581499
  review:
    summary: >-
      Interactions with CDC20 and MAD1L1 from study of nuclear pore-mediated anaphase
      inhibitor assembly.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere.
    supported_by:
    - reference_id: PMID:24581499
      supporting_text: "Nuclear pores protect genome integrity by assembling a premitotic
        and Mad1-dependent anaphase inhibitor"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25383541
  review:
    summary: >-
      Interactions with BUB1B and CDC20 from study of MCC binding to second CDC20.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core MCC interactions captured elsewhere.
    supported_by:
    - reference_id: PMID:25383541
      supporting_text: "The mitotic checkpoint complex binds a second CDC20 to inhibit
        active APC/C"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25416956
  review:
    summary: >-
      Multiple interactions (SDCBP, KEAP1, p31comet, TSC22D4) from proteome-scale
      interactome map.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      HTP interactome study. Core interactions captured elsewhere; peripheral
      interactions not clearly related to checkpoint function.
    supported_by:
    - reference_id: PMID:25416956
      supporting_text: "A proteome-scale map of the human interactome network"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25502805
  review:
    summary: >-
      Interaction with p31comet from disease mutation phenotyping study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      p31comet interaction captured elsewhere.
    supported_by:
    - reference_id: PMID:25502805
      supporting_text: "A massively parallel pipeline to clone DNA variants and examine
        molecular phenotypes of human disease mutations"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25852190
  review:
    summary: >-
      Interaction with BUB1B from kinase network analysis in TRAIL-induced apoptosis.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      BUB1B/BUBR1 interaction captured in MCC annotations.
    supported_by:
    - reference_id: PMID:25852190
      supporting_text: "Integrative analysis of kinase networks in TRAIL-induced apoptosis
        provides a source of potential targets for combination therapy"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:26258632
  review:
    summary: >-
      Interaction with MAD1L1 from study of MAD1 role in chromosome congression.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations.
    supported_by:
    - reference_id: PMID:26258632
      supporting_text: "Mad1 promotes chromosome congression by anchoring a kinesin
        motor to the kinetochore"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:26496610
  review:
    summary: >-
      Multiple interactions (CDC27, CDC20, p31comet, MAD1L1) from quantitative
      interactome study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere. HTP study with stoichiometry data.
    supported_by:
    - reference_id: PMID:26496610
      supporting_text: "A human interactome in three quantitative dimensions organized
        by stoichiometries and abundances"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:28514442
  review:
    summary: >-
      Interaction with TSC22D4 from disease network study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      TSC22D4 interaction not clearly related to checkpoint function. HTP study.
    supported_by:
    - reference_id: PMID:28514442
      supporting_text: "Architecture of the human interactome defines protein communities
        and disease networks"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:29997244
  review:
    summary: >-
      Interaction with MAD1L1 from bioluminescence two-hybrid study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations.
    supported_by:
    - reference_id: PMID:29997244
      supporting_text: "LuTHy: a double-readout bioluminescence-based two-hybrid technology
        for quantitative mapping of protein-protein interactions"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:31467278
  review:
    summary: >-
      Interaction with MAD1L1 from binary interactome mapping study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations. HTP study.
    supported_by:
    - reference_id: PMID:31467278
      supporting_text: "Maximizing binary interactome mapping with a minimal number
        of assays"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:31515488
  review:
    summary: >-
      Interactions with KEAP1, p31comet, MAD1L1 from variant disruption study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere. Study addresses variant effects on interactions.
    supported_by:
    - reference_id: PMID:31515488
      supporting_text: "Extensive disruption of protein interactions by genetic variants
        across the allele frequency spectrum in human populations"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:32296183
  review:
    summary: >-
      Multiple interactions (DEPDC5, INSR, KEAP1, p31comet, EPM2AIP1, MAD1L1) from
      human binary interactome reference map.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere. Several peripheral interactions (DEPDC5,
      INSR, EPM2AIP1) not clearly related to checkpoint function.
    supported_by:
    - reference_id: PMID:32296183
      supporting_text: "A reference map of the human binary protein interactome"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:32814053
  review:
    summary: >-
      Interaction with MAD1L1 from neurodegenerative disease interactome study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations.
    supported_by:
    - reference_id: PMID:32814053
      supporting_text: "Interactome Mapping Provides a Network of Neurodegenerative
        Disease Proteins and Uncovers Widespread Protein Aggregation"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:33961781
  review:
    summary: >-
      Multiple interactions (BUB1B, INSR, CDC27, CDC20, p31comet, SGO2, EPM2AIP1,
      TSC22D4, MAD1L1) from cell-specific interactome study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere. HTP dual proteome-scale study.
    supported_by:
    - reference_id: PMID:33961781
      supporting_text: "Dual proteome-scale networks reveal cell-specific remodeling
        of the human interactome"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:35384245
  review:
    summary: >-
      Interaction with INSR (insulin receptor) from RTK interactome study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      INSR interaction not clearly related to checkpoint function. May represent
      regulatory cross-talk or indirect interaction.
    supported_by:
    - reference_id: PMID:35384245
      supporting_text: "Physical and functional interactome atlas of human receptor
        tyrosine kinases"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:37398436
  review:
    summary: >-
      Interaction with MAD1L1 from AI-guided PPI drug discovery study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations.
    supported_by:
    - reference_id: PMID:37398436
      supporting_text: "AI-guided pipeline for protein-protein interaction drug discovery
        identifies a SARS-CoV-2 inhibitor"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:37926298
  review:
    summary: >-
      Interaction with CDC20 from colorectal cancer therapeutic study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core CDC20 interaction captured elsewhere.
    supported_by:
    - reference_id: PMID:37926298
      supporting_text: "Therapeutic role of 2-stearoxyphenethyl phosphocholine targeting
        microtubule dynamics"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:40205054
  review:
    summary: >-
      Multiple interactions (BUB1B, CDC27, CDC20, p31comet, EPM2AIP1, MAD1L1) from
      multimodal cell maps study.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core interactions captured elsewhere. HTP multimodal study.
    supported_by:
    - reference_id: PMID:40205054
      supporting_text: "Multimodal cell maps as a foundation for structural and functional
        genomics"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:9092546
  review:
    summary: >-
      Interaction with insulin receptor (INSR) documented. MAD2 interacts with insulin
      receptor C-terminus but not IGF1R.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Interaction with insulin receptor is not clearly related to core checkpoint
      function.
      May represent regulatory cross-talk between cell cycle and insulin signaling.
    supported_by:
    - reference_id: PMID:9092546
      supporting_text: "Interaction of MAD2 with the carboxyl terminus of the insulin
        receptor but not with the IGFIR"

- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:16525508
  review:
    summary: >-
      MAD2 homodimerization demonstrated. MAD2 forms conformational heterodimers
      between
      O-MAD2 and C-MAD2 forms.
    action: ACCEPT
    reason: >-
      MAD2 homodimerization (O-MAD2:C-MAD2) is essential for the template model
      of
      MAD2 activation at kinetochores. This is a core molecular function.
    supported_by:
    - reference_id: PMID:18022367
      supporting_text: "Third, O-Mad2 and C-Mad2 engage in a \"conformational\" dimer
        that is essential for spindle checkpoint function in different organisms"
    - reference_id: PMID:16525508
      supporting_text: "Determinants of conformational dimerization of Mad2 and its
        inhibition by p31comet"

- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:18022367
  review:
    summary: >-
      Crystal structure of O-MAD2:C-MAD2 conformational dimer reported. Essential
      for
      checkpoint function.
    action: ACCEPT
    reason: >-
      Landmark structural study establishing the molecular basis of MAD2 conformational
      dimerization. Core molecular function for SAC signaling.
    supported_by:
    - reference_id: PMID:18022367
      supporting_text: "The crystal structure of the O-Mad2-C-Mad2 conformational
        dimer...reveals an asymmetric interface that explains the selective dimerization
        of the O-Mad2 and C-Mad2 conformers"

- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:18318601
  review:
    summary: >-
      Crystal structure of symmetric C-MAD2:C-MAD2 dimer reported. Provides insights
      into MAD2 regulation.
    action: ACCEPT
    reason: >-
      Demonstrates both asymmetric (O-MAD2:C-MAD2) and symmetric (C-MAD2:C-MAD2)
      dimerization modes. Relevant to understanding MAD2 regulation.
    supported_by:
    - reference_id: PMID:18318601
      supporting_text: "Insights into mad2 regulation in the spindle checkpoint revealed
        by the crystal structure of the symmetric mad2 dimer"

- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:21041666
  review:
    summary: >-
      MAD2 homodimerization studied in context of phosphorylation regulation.
    action: ACCEPT
    reason: >-
      Confirms homodimerization as functionally important and phosphoregulated.
    supported_by:
    - reference_id: PMID:21041666
      supporting_text: "Phosphorylation of the spindle checkpoint protein Mad2 regulates
        its conformational transition"

- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:21772247
  review:
    summary: >-
      MAD2 homodimerization confirmed in in vivo functional study.
    action: ACCEPT
    reason: >-
      In vivo validation of MAD2 homodimerization function.
    supported_by:
    - reference_id: PMID:21772247
      supporting_text: "Probing the in vivo function of Mad1:C-Mad2 in the spindle
        assembly checkpoint"

- term:
    id: GO:0000775
    label: chromosome, centromeric region
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  review:
    summary: >-
      Ensembl Compara transfer from mouse ortholog. MAD2 localizes to centromeric
      regions via kinetochore association.
    action: ACCEPT
    reason: >-
      Kinetochores are located at centromeric regions. This annotation is consistent
      with MAD2 kinetochore localization.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "Human, or Homo sapiens, MAD2 (hsMAD2) was localized at the
        kinetochore after chromosome condensation"

- term:
    id: GO:0005694
    label: chromosome
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  review:
    summary: >-
      Ensembl Compara transfer from mouse ortholog. MAD2 associates with chromosomes
      via kinetochore localization.
    action: ACCEPT
    reason: >-
      Broader term encompassing kinetochore/centromere localization. Accurate but
      less specific than kinetochore annotation.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "Human, or Homo sapiens, MAD2 (hsMAD2) was localized at the
        kinetochore after chromosome condensation"

- term:
    id: GO:0007094
    label: mitotic spindle assembly checkpoint signaling
  evidence_type: NAS
  original_reference_id: PMID:11535616
  review:
    summary: >-
      Non-traceable author statement from study of checkpoint inhibition of APC/C
      in HeLa cells. MAD2 is part of the MCC that inhibits APC/C.
    action: ACCEPT
    reason: >-
      Core biological process for MAD2. This study describes MCC (BUBR1, BUB3, CDC20,
      MAD2) mediated APC/C inhibition.
    supported_by:
    - reference_id: PMID:11535616
      supporting_text: "Checkpoint inhibition of the APC/C in HeLa cells is mediated
        by a complex of BUBR1, BUB3, CDC20, and MAD2"

- term:
    id: GO:0007094
    label: mitotic spindle assembly checkpoint signaling
  evidence_type: IMP
  original_reference_id: PMID:23509069
  review:
    summary: >-
      Mutant phenotype evidence from MISP (mitotic spindle positioning) study.
      MAD2 involvement in spindle checkpoint demonstrated through genetic perturbation.
    action: ACCEPT
    reason: >-
      IMP evidence confirms MAD2 role in SAC signaling. Core biological process.
    supported_by:
    - reference_id: PMID:23509069
      supporting_text: "MISP is a novel Plk1 substrate required for proper spindle
        orientation and mitotic progression"

- term:
    id: GO:0090267
    label: positive regulation of mitotic cell cycle spindle assembly checkpoint
  evidence_type: IDA
  original_reference_id: PMID:22898774
  review:
    summary: >-
      Direct assay evidence from review of mitotic checkpoint evolution and function.
      MAD2 positively regulates the SAC.
    action: ACCEPT
    reason: >-
      MAD2 activation (O-MAD2 to C-MAD2 conversion) is essential for SAC activation.
      This is a core regulatory function.
    supported_by:
    - reference_id: PMID:22898774
      supporting_text: "Evolution and function of the mitotic checkpoint"

- term:
    id: GO:0090267
    label: positive regulation of mitotic cell cycle spindle assembly checkpoint
  evidence_type: IDA
  original_reference_id: PMID:8824189
  review:
    summary: >-
      Original discovery paper demonstrating MAD2 is necessary for mitotic checkpoint
      function in HeLa cells.
    action: ACCEPT
    reason: >-
      Foundational paper establishing MAD2 as essential for SAC function. Antibody
      electroporation experiments directly demonstrated MAD2 is required for checkpoint.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "The human homolog of MAD2 was isolated and shown to be a necessary
        component of the mitotic checkpoint in HeLa cells by antibody electroporation
        experiments"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19010891
  review:
    summary: >-
      Interaction with MAD1L1 isoforms demonstrated. MAD1beta sequesters MAD2 in
      cytoplasm.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      MAD1-MAD2 interaction captured in specific annotations. This study addresses
      a cancer-associated MAD1 splice variant.
    supported_by:
    - reference_id: PMID:19010891
      supporting_text: "MAD1beta was found to physically interact with MAD2 and sequester
        it in the cytoplasm"

- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IDA
  original_reference_id: PMID:19010891
  review:
    summary: >-
      Direct assay showing nuclear localization. MAD1alpha is in nucleus while MAD1beta
      is cytoplasmic, affecting MAD2 localization.
    action: ACCEPT
    reason: >-
      Nuclear localization is validated by multiple studies. MAD2 is recruited to
      nuclear pore complex by TPR during interphase.
    supported_by:
    - reference_id: PMID:19010891
      supporting_text: "MAD1alpha was found in the nucleus"
    - reference_id: PMID:18981471
      supporting_text: "Depletion of Tpr in HeLa cells disrupts the NPC localization
        of Mad1 and Mad2 during interphase"

- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IDA
  original_reference_id: PMID:8824189
  review:
    summary: >-
      Original discovery paper showing MAD2 kinetochore localization. MAD2 localizes
      to kinetochores after chromosome condensation but not at metaphase.
    action: ACCEPT
    reason: >-
      Foundational evidence for MAD2 kinetochore localization. This is core to
      checkpoint signaling mechanism.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "Human, or Homo sapiens, MAD2 (hsMAD2) was localized at the
        kinetochore after chromosome condensation but was no longer observed at the
        kinetochore in metaphase, suggesting that MAD2 might monitor the completeness
        of the spindle-kinetochore attachment"

- term:
    id: GO:0007094
    label: mitotic spindle assembly checkpoint signaling
  evidence_type: IDA
  original_reference_id: PMID:18981471
  review:
    summary: >-
      Direct evidence showing TPR binds MAD1 and MAD2 and is important for SAC
      signaling. MAD2 activation by MAD1 is required for APC-CDC20 inhibition.
    action: ACCEPT
    reason: >-
      Key study establishing TPR role in MAD1-MAD2 regulation. Confirms MAD2
      essential role in SAC signaling.
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "Depletion of Tpr decreases the levels of Mad1 at kinetochores
        during prometaphase, correlating with the inability of Mad1 to activate Mad2,
        which is required for inhibiting APC(Cdc20)"

- term:
    id: GO:0044615
    label: nuclear pore nuclear basket
  evidence_type: IDA
  original_reference_id: PMID:18981471
  review:
    summary: >-
      Direct evidence showing MAD2 localizes to nuclear pore complex via TPR
      during interphase.
    action: ACCEPT
    reason: >-
      Important localization for early MCC assembly. TPR is a nuclear pore basket
      component that recruits MAD1-MAD2 during interphase.
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "Depletion of Tpr in HeLa cells disrupts the NPC localization
        of Mad1 and Mad2 during interphase"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25422469
  review:
    summary: >-
      Interaction with FAT10/UBD (O15205) demonstrated. Disrupting FAT10-MAD2
      binding inhibits tumor progression.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      UBD/FAT10 interaction may regulate MAD2 during mitosis but is not part of
      core checkpoint mechanism. Represents regulatory/peripheral function.
    supported_by:
    - reference_id: PMID:25422469
      supporting_text: "Disruption of FAT10-MAD2 binding inhibits tumor progression"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:18794143
  review:
    summary: >-
      Interaction with HSF1 (Q00613) demonstrated. HSF1 interacts with MAD2 during
      mitosis.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      HSF1 interaction occurs during mitosis but is not part of core SAC mechanism.
      May represent regulatory cross-talk between stress response and cell cycle.
    supported_by:
    - reference_id: PMID:18794143
      supporting_text: "HSF1 as a mitotic regulator: phosphorylation of HSF1 by Plk1
        is essential for mitotic progression"

- term:
    id: GO:0045930
    label: negative regulation of mitotic cell cycle
  evidence_type: IMP
  original_reference_id: PMID:21274008
  review:
    summary: >-
      Mutant phenotype evidence from study of MTBP role in mitotic progression.
      MAD2 negatively regulates mitotic cell cycle by preventing premature anaphase.
    action: ACCEPT
    reason: >-
      MAD2-mediated SAC activation delays metaphase-to-anaphase transition, thereby
      negatively regulating mitotic cell cycle progression.
    supported_by:
    - reference_id: PMID:21274008
      supporting_text: "MTBP plays a crucial role in mitotic progression and chromosome
        segregation"

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141409
  review:
    summary: >-
      Reactome annotation for MAD1 binding to kinetochore. MAD2 is present in cytosol
      before activation.
    action: ACCEPT
    reason: >-
      Cytosolic O-MAD2 pool is essential for checkpoint activation. Reactome pathway
      correctly places MAD2 in cytosol before kinetochore recruitment.
    supported_by:
    - reference_id: PMID:18022367
      supporting_text: "a catalytic model in which a C-Mad2 template facilitates the
        binding of O-Mad2 to Cdc20"

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141422
  review:
    summary: >-
      Reactome annotation for MAD2 conversion to inhibitory state via MAD1 interaction.
    action: ACCEPT
    reason: >-
      Cytosolic localization is prerequisite for MAD2 recruitment and activation.
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "The mitotic arrest-deficient protein Mad1 forms a complex
        with Mad2"

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141431
  review:
    summary: >-
      Reactome annotation for MAD2 association with MAD1 kinetochore complex.
    action: ACCEPT
    reason: >-
      Consistent with template model of MAD2 activation.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141439
  review:
    summary: >-
      Reactome annotation for release of activated MAD2 from kinetochores.
    action: ACCEPT
    reason: >-
      Activated C-MAD2:CDC20 is released to cytosol to form MCC.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-1638803
  review:
    summary: >-
      Reactome annotation for PLK1 phosphorylation of cohesin at centromeres.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in cohesin regulation is secondary to core checkpoint function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-1638821
  review:
    summary: >-
      Reactome annotation for PP2A-B56 dephosphorylation of centromeric cohesin.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in cohesin regulation is secondary to core checkpoint function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-2467809
  review:
    summary: >-
      Reactome annotation for ESPL1 (Separase) cleavage of centromeric cohesin.
    action: ACCEPT
    reason: >-
      Cytosolic localization during sister chromatid separation.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-2467811
  review:
    summary: >-
      Reactome annotation for separation of sister chromatids.
    action: ACCEPT
    reason: >-
      Cytosolic localization during sister chromatid separation.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-2468287
  review:
    summary: >-
      Reactome annotation for CDK1 phosphorylation of CDCA5 (Sororin) at centromeres.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in sororin regulation is secondary to core checkpoint function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-2484822
  review:
    summary: >-
      Reactome annotation for kinetochore assembly.
    action: ACCEPT
    reason: >-
      Cytosolic MAD2 is recruited during kinetochore assembly.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-375302
  review:
    summary: >-
      Reactome annotation for kinetochore capture of astral microtubules.
    action: ACCEPT
    reason: >-
      Cytosolic localization during spindle assembly.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-5666129
  review:
    summary: >-
      Reactome annotation for CDC42:GTP recruiting DIAPH2-2 to kinetochores.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in DIAPH2 recruitment is secondary to core checkpoint function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-5666160
  review:
    summary: >-
      Reactome annotation for AURKB phosphorylation of DIAPH2-2 at kinetochores.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in DIAPH2 phosphorylation is secondary to core checkpoint
      function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-5666169
  review:
    summary: >-
      Reactome annotation for kinetochore capture regulation by CDC42:GTP:p-DIAPH2-2.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in this process is secondary to core checkpoint function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-9648114
  review:
    summary: >-
      Reactome annotation for EML4 recruiting NUDC to mitotic spindle.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 involvement in EML4-NUDC pathway is secondary to core checkpoint function.

- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IDA
  original_reference_id: PMID:20133940
  review:
    summary: >-
      Direct evidence for MAD2 kinetochore localization from TPR-dynein study.
    action: ACCEPT
    reason: >-
      Additional experimental validation of MAD2 kinetochore localization.
    supported_by:
    - reference_id: PMID:20133940
      supporting_text: "Tpr orchestrates proper chromosome segregation through interaction
        with dynein light chain...spindle checkpoints Mad1 and Mad2"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:20133940
  review:
    summary: >-
      Interaction with TPR (P12270) demonstrated.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      TPR interaction is important for MAD2 NPC localization but "protein binding"
      is too vague. Functional consequence is captured in localization annotations.
    supported_by:
    - reference_id: PMID:20133940
      supporting_text: "Here, we show association of another nucleoporin, termed Tpr
        (translocated promoter region), with the molecular motors dynein and dynactin,
        which both orchestrate with the spindle checkpoints Mad1 and Mad2 during cell
        division"

- term:
    id: GO:0072686
    label: mitotic spindle
  evidence_type: IDA
  original_reference_id: PMID:20133940
  review:
    summary: >-
      Direct evidence for MAD2 localization to mitotic spindle.
    action: ACCEPT
    reason: >-
      Mitotic spindle localization is consistent with MAD2 role in SAC signaling.
    supported_by:
    - reference_id: PMID:20133940
      supporting_text: "Here, we show association of another nucleoporin, termed Tpr
        (translocated promoter region), with the molecular motors dynein and dynactin"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19273613
  review:
    summary: >-
      Interaction with TPR (Megator ortholog) from study of spindle matrix protein.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      TPR interaction captured elsewhere.
    supported_by:
    - reference_id: PMID:19273613
      supporting_text: "Spatiotemporal control of mitosis by the conserved spindle
        matrix protein Megator"

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141423
  review:
    summary: >-
      Reactome annotation for MCC complex binding to APC/C complex.
    action: ACCEPT
    reason: >-
      Core checkpoint mechanism. MCC assembly and APC/C inhibition occur in cytosol.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141429
  review:
    summary: >-
      Reactome annotation for APC/C inactivation via CDC20 sequestration.
    action: ACCEPT
    reason: >-
      Core checkpoint mechanism. CDC20 sequestration by MCC occurs in cytosol.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-141437
  review:
    summary: >-
      Reactome annotation for MCC complex formation.
    action: ACCEPT
    reason: >-
      Core checkpoint mechanism. MCC (MAD2, BUBR1, BUB3, CDC20) forms in cytosol.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-174104
  review:
    summary: >-
      Reactome annotation for ubiquitination of Cyclin A by APC/C:Cdc20.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 in MCC inhibits this reaction. Secondary to core checkpoint signaling.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-174171
  review:
    summary: >-
      Reactome annotation for Cyclin A association with APC/C.
    action: KEEP_AS_NON_CORE
    reason: >-
      Secondary pathway annotation.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-174238
  review:
    summary: >-
      Reactome annotation for APC/C:Cdc20 activation by MCC dissociation.
    action: ACCEPT
    reason: >-
      Describes checkpoint silencing when MCC dissociates.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-174255
  review:
    summary: >-
      Reactome annotation for multiubiquitinated Cyclin A degradation.
    action: KEEP_AS_NON_CORE
    reason: >-
      Downstream consequence of checkpoint silencing.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-179410
  review:
    summary: >-
      Reactome annotation for Nek2A association with MCC:APC/C.
    action: KEEP_AS_NON_CORE
    reason: >-
      MAD2 as part of MCC is involved but this is secondary function.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-179417
  review:
    summary: >-
      Reactome annotation for Nek2A multiubiquitination.
    action: KEEP_AS_NON_CORE
    reason: >-
      Secondary pathway annotation.

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-179421
  review:
    summary: >-
      Reactome annotation for Nek2A degradation.
    action: KEEP_AS_NON_CORE
    reason: >-
      Secondary pathway annotation.

- term:
    id: GO:1904667
    label: negative regulation of ubiquitin protein ligase activity
  evidence_type: IDA
  original_reference_id: PMID:11459825
  review:
    summary: >-
      Direct evidence that MAD2-related protein inhibits APC/C ubiquitin ligase
      activity. Note: This paper primarily studies MAD2L2, not MAD2L1, but
      proposes mechanism for MAD2 inhibition of APC/C.
    action: ACCEPT
    reason: >-
      MAD2 inhibits APC/C-CDC20 ubiquitin ligase activity through MCC formation.
      This is a core molecular function of MAD2 in the SAC.
    supported_by:
    - reference_id: PMID:11459825
      supporting_text: "We suggest that MAD2L2 and MAD2 inhibit the release of substrates
        from APC and propose a mechanism of inhibition"
    - reference_id: PMID:10700282
      supporting_text: "The checkpoint protein Mad2 inhibits the activity of the anaphase
        promoting complex"

- term:
    id: GO:0042177
    label: negative regulation of protein catabolic process
  evidence_type: IDA
  original_reference_id: PMID:11459825
  review:
    summary: >-
      By inhibiting APC/C, MAD2 prevents degradation of APC/C substrates like
      cyclins and securin.
    action: ACCEPT
    reason: >-
      Accurate annotation. MAD2-mediated MCC inhibition of APC/C prevents
      ubiquitination and subsequent proteasomal degradation of mitotic substrates.
    supported_by:
    - reference_id: PMID:11459825
      supporting_text: "We suggest that MAD2L2 and MAD2 inhibit the release of substrates
        from APC"

- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IDA
  original_reference_id: PMID:20870947
  review:
    summary: >-
      Direct evidence for nuclear localization from Pcid2 study in B cells.
    action: ACCEPT
    reason: >-
      Additional experimental validation of nuclear localization.
    supported_by:
    - reference_id: PMID:20870947
      supporting_text: "Critical role of Pcid2 in B cell survival through the regulation
        of MAD2 expression"

- term:
    id: GO:0048471
    label: perinuclear region of cytoplasm
  evidence_type: IDA
  original_reference_id: PMID:20870947
  review:
    summary: >-
      Direct evidence for perinuclear localization from Pcid2 study.
    action: ACCEPT
    reason: >-
      Perinuclear localization is consistent with MAD2 association with nuclear
      pore complex.
    supported_by:
    - reference_id: PMID:20870947
      supporting_text: "Critical role of Pcid2 in B cell survival through the regulation
        of MAD2 expression"

- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IDA
  original_reference_id: PMID:19229290
  review:
    summary: >-
      Direct evidence for kinetochore localization from dynein study.
    action: ACCEPT
    reason: >-
      Additional experimental validation of kinetochore localization.
    supported_by:
    - reference_id: PMID:19229290
      supporting_text: "Dynein light intermediate chain 1 is required for progress
        through the spindle assembly checkpoint"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:10200259
  review:
    summary: >-
      Interaction with FAT10/UBD (O15205) demonstrated. FAT10 binds MAD2 non-covalently.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      FAT10 interaction may regulate MAD2 but is not core checkpoint mechanism.
    supported_by:
    - reference_id: PMID:10200259
      supporting_text: "A MHC-encoded ubiquitin-like protein (FAT10) binds noncovalently
        to the spindle assembly checkpoint protein MAD2"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:15525512
  review:
    summary: >-
      Interaction with CDC20 documented. Study of BUB1 phosphorylation of CDC20.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Core CDC20 interaction captured in more specific annotations.
    supported_by:
    - reference_id: PMID:15525512
      supporting_text: "Phosphorylation of Cdc20 by Bub1 provides a catalytic mechanism
        for APC/C inhibition by the spindle checkpoint"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:10700282
  review:
    summary: >-
      Seminal structural study showing MAD2-CDC20 interaction. MAD2 C-terminal
      region binds CDC20.
    action: MODIFY
    reason: >-
      This is the primary MAD2-CDC20 interaction paper. Should be annotated to
      more specific complex term.
    proposed_replacement_terms:
    - id: GO:1990333
      label: mitotic checkpoint complex, CDC20-MAD2 subcomplex
    supported_by:
    - reference_id: PMID:10700282
      supporting_text: "Mad2 and Cdc20 form a tight 1:1 heterodimeric complex in which
        the C-terminal segment of Mad2 becomes folded"

- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IDA
  original_reference_id: PMID:19468067
  review:
    summary: >-
      Colocalization with kinetochore markers from Spindly study.
    action: ACCEPT
    reason: >-
      Additional experimental validation of kinetochore localization.
    supported_by:
    - reference_id: PMID:19468067
      supporting_text: "Mitotic control of kinetochore-associated dynein and spindle
        orientation by human Spindly"

- term:
    id: GO:0000776
    label: kinetochore
  evidence_type: IDA
  original_reference_id: PMID:17363900
  review:
    summary: >-
      Colocalization evidence from Nup107-160 nuclear pore subcomplex study.
    action: ACCEPT
    reason: >-
      Additional experimental validation of kinetochore localization.
    supported_by:
    - reference_id: PMID:17363900
      supporting_text: "The human Nup107-160 nuclear pore subcomplex contributes to
        proper kinetochore functions"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:18981471
  review:
    summary: >-
      Interaction with TPR (P12270) demonstrated. TPR directly binds MAD1 and MAD2.
    action: MODIFY
    reason: >-
      TPR interaction is functionally important for MAD2 NPC localization during
      interphase. Should be captured with more specific term if available.
    proposed_replacement_terms:
    - id: GO:0044615
      label: nuclear pore nuclear basket
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "Tpr directly binds to Mad1 and Mad2"

- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IDA
  original_reference_id: PMID:18981471
  review:
    summary: >-
      Direct evidence for nuclear localization via TPR association.
    action: ACCEPT
    reason: >-
      Key study establishing MAD2 NPC localization during interphase.
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "Depletion of Tpr in HeLa cells disrupts the NPC localization
        of Mad1 and Mad2 during interphase"

- term:
    id: GO:0005829
    label: cytosol
  evidence_type: IDA
  original_reference_id: PMID:18981471
  review:
    summary: >-
      Direct evidence for cytosolic localization. O-MAD2 pool is cytosolic.
    action: ACCEPT
    reason: >-
      Cytosolic localization is essential for template model of MAD2 activation.
    supported_by:
    - reference_id: PMID:18981471
      supporting_text: "Tpr directly binds to Mad1 and Mad2...decreases the levels
        of Mad1-bound Mad2"

- term:
    id: GO:0042803
    label: protein homodimerization activity
  evidence_type: IPI
  original_reference_id: PMID:18022367
  review:
    summary: >-
      Seminal structural study demonstrating MAD2 homodimerization. O-MAD2:C-MAD2
      conformational dimer is essential for checkpoint function.
    action: ACCEPT
    reason: >-
      This is a core molecular function of MAD2. The conformational dimer enables
      template-based activation of O-MAD2 to C-MAD2.
    supported_by:
    - reference_id: PMID:18022367
      supporting_text: "Third, O-Mad2 and C-Mad2 engage in a \"conformational\" dimer
        that is essential for spindle checkpoint function in different organisms"

- term:
    id: GO:0048471
    label: perinuclear region of cytoplasm
  evidence_type: IDA
  original_reference_id: PMID:8824189
  review:
    summary: >-
      Original discovery paper noting MAD2 perinuclear localization.
    action: ACCEPT
    reason: >-
      Perinuclear localization consistent with NPC association.
    supported_by:
    - reference_id: PMID:8824189
      supporting_text: "Identification of a human mitotic checkpoint gene: hsMAD2"

- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:12006501
  review:
    summary: >-
      Interaction with MAD1L1 demonstrated in structural study of tetrameric
      Mad1-Mad2 core complex.
    action: MODIFY
    reason: >-
      This is a core functional interaction. The MAD1-MAD2 complex is essential
      for checkpoint signaling. Should be annotated to specific complex term.
    proposed_replacement_terms:
    - id: GO:1990728
      label: mitotic spindle assembly checkpoint MAD1-MAD2 complex
    supported_by:
    - reference_id: PMID:12006501
      supporting_text: "The crystal structure of the Mad1-Mad2 complex reveals an
        asymmetric tetramer, with elongated Mad1 monomers parting from a coiled-coil
        to form two connected sub-complexes with Mad2"

# Add annotation for MCC membership
- term:
    id: GO:0033597
    label: mitotic checkpoint complex
  evidence_type: IDA
  original_reference_id: PMID:11535616
  review:
    summary: >-
      MAD2 is a core component of the mitotic checkpoint complex (MCC) which consists
      of MAD2, BUBR1/MAD3, BUB3, and CDC20 in mammals.
    action: NEW
    reason: >-
      The MCC annotation should be added as it represents a core complex for MAD2
      function in SAC signaling. The MCC inhibits APC/C-CDC20 to prevent premature
      anaphase.
    supported_by:
    - reference_id: PMID:11535616
      supporting_text: "Checkpoint inhibition of the APC/C in HeLa cells is mediated
        by a complex of BUBR1, BUB3, CDC20, and MAD2"
    - reference_id: PMID:10700282
      supporting_text: "The checkpoint protein Mad2 inhibits the activity of the anaphase
        promoting complex by sequestering Cdc20"

- term:
    id: GO:1990948
    label: ubiquitin ligase inhibitor activity
  evidence_type: NAS
  review:
    summary: Added to align core_functions with existing annotations.
    action: NEW
    reason: Core function term not present in existing_annotations.
    supported_by:
    - reference_id: PMID:10700282
      supporting_text: "The checkpoint protein Mad2 inhibits the activity of the anaphase
        promoting complex by sequestering Cdc20"
    - reference_id: PMID:11459825
      supporting_text: "We suggest that MAD2L2 and MAD2 inhibit the release of substrates
        from APC"

references:
- id: GO_REF:0000033
  title: Annotation inferences using phylogenetic trees
  findings: []
- id: GO_REF:0000043
  title: Gene Ontology annotation based on UniProtKB/Swiss-Prot keyword mapping
  findings: []
- id: GO_REF:0000107
  title: Automatic transfer of experimentally verified manual GO annotation data
    to orthologs using Ensembl Compara
  findings: []
- id: GO_REF:0000117
  title: Electronic Gene Ontology annotations created by ARBA machine learning
    models
  findings: []
- id: GO_REF:0000120
  title: Combined Automated Annotation using Multiple IEA Methods
  findings: []
- id: PMID:8824189
  title: 'Identification of a human mitotic checkpoint gene: hsMAD2.'
  findings:
  - statement: MAD2 is localized at kinetochores after chromosome condensation
      but absent at metaphase
  - statement: MAD2 is required for mitotic checkpoint function in HeLa cells
- id: PMID:10700282
  title: Structure of the Mad2 spindle assembly checkpoint protein and its
    interaction with Cdc20.
  findings:
  - statement: MAD2 forms tight 1:1 complex with CDC20
  - statement: C-terminal region of MAD2 is required for CDC20 binding
- id: PMID:11459825
  title: 'Inhibition of Cdh1-APC by the MAD2-related protein MAD2L2: a novel mechanism
    for regulating Cdh1.'
  findings:
  - statement: MAD2 family proteins inhibit APC ubiquitin ligase activity
- id: PMID:11535616
  title: Checkpoint inhibition of the APC/C in HeLa cells is mediated by a
    complex of BUBR1, BUB3, CDC20, and MAD2.
  findings:
  - statement: MCC (BUBR1, BUB3, CDC20, MAD2) mediates checkpoint inhibition of
      APC/C
- id: PMID:12006501
  title: 'Crystal structure of the tetrameric Mad1-Mad2 core complex: implications of a ''safety belt'' binding mechanism for the spindle checkpoint.'
  findings:
  - statement: MAD1-MAD2 forms asymmetric tetramer
  - statement: Safety belt binding mechanism for ligand binding
- id: PMID:18022367
  title: "The Mad2 conformational dimer: structure and implications for the spindle assembly checkpoint."
  findings:
  - statement: O-MAD2 and C-MAD2 form conformational heterodimer
  - statement: Essential for checkpoint function
- id: PMID:18022368
  title: p31comet blocks Mad2 activation through structural mimicry
  findings:
  - statement: p31comet binds MAD2 and blocks its activation
  - statement: p31comet is structurally similar to MAD2
- id: PMID:18981471
  title: "Tpr directly binds to Mad1 and Mad2 and is important for the Mad1-Mad2-mediated mitotic spindle checkpoint."
  findings:
  - statement: TPR recruits MAD1-MAD2 to nuclear pore complex during interphase
  - statement: TPR depletion disrupts checkpoint signaling
- id: PMID:10527948
  title: Evidence for an interaction of the metalloprotease-disintegrin tumour
    necrosis factor alpha convertase (TACE) with mitotic arrest deficient 2
    (MAD2), and of the metalloprotease-disintegrin MDC9 with a novel
    MAD2-related protein, MAD2beta.
  findings: []
- id: PMID:15182668
  title: Spindle checkpoint protein dynamics at kinetochores in living cells.
  findings: []
- id: PMID:16189514
  title: Towards a proteome-scale map of the human protein-protein interaction
    network
  findings: []
- id: PMID:16525508
  title: Determinants of conformational dimerization of Mad2 and its inhibition
    by p31comet
  findings:
  - statement: MAD2 conformational dimerization is essential for checkpoint
      function
- id: PMID:17443180
  title: Anaphase initiation is regulated by antagonistic ubiquitination and
    deubiquitination activities
  findings: []
- id: PMID:17443186
  title: Ubiquitination by the anaphase-promoting complex drives spindle
    checkpoint inactivation
  findings: []
- id: PMID:18318601
  title: "Insights into mad2 regulation in the spindle checkpoint revealed by the crystal structure of the symmetric mad2 dimer."
  findings:
  - statement: MAD2 can form symmetric C-MAD2:C-MAD2 dimer
- id: PMID:18692475
  title: A protein domain-based interactome network for C. elegans early
    embryogenesis
  findings: []
- id: PMID:19143472
  title: The influence of catalysis on Mad2 activation dynamics
  findings: []
- id: PMID:19615732
  title: Defining the human deubiquitinating enzyme interaction landscape
  findings: []
- id: PMID:20212161
  title: ATP is required for the release of the anaphase-promoting
    complex/cyclosome from inhibition by the mitotic checkpoint.
  findings: []
- id: PMID:20360068
  title: Systematic analysis of human protein complexes identifies chromosome
    segregation proteins
  findings: []
- id: PMID:20951947
  title: Pharmacologic inhibition of the anaphase-promoting complex induces a
    spindle checkpoint-dependent mitotic arrest in the absence of spindle
    damage.
  findings: []
- id: PMID:21041666
  title: Phosphorylation of the spindle checkpoint protein Mad2 regulates its
    conformational transition
  findings:
  - statement: MAD2 phosphorylation regulates conformational switching
- id: PMID:21300909
  title: p31comet promotes disassembly of the mitotic checkpoint complex in an
    ATP-dependent process
  findings:
  - statement: p31comet and TRIP13 promote MCC disassembly
- id: PMID:21407176
  title: "Evidence that Aurora B is implicated in spindle checkpoint signalling independently of error correction."
  findings: []
- id: PMID:21666598
  title: Shugoshin is a Mad1/Cdc20-like interactor of Mad2
  findings:
  - statement: SGO2 binds MAD2 similarly to MAD1 and CDC20
- id: PMID:21772247
  title: Probing the in vivo function of Mad1:C-Mad2 in the spindle assembly
    checkpoint
  findings: []
- id: PMID:21988832
  title: Toward an understanding of the protein interaction network of the human
    liver
  findings: []
- id: PMID:22000412
  title: Structure of a Blinkin-BUBR1 complex reveals an interaction crucial for
    kinetochore-mitotic checkpoint regulation via an unanticipated binding Site.
  findings: []
- id: PMID:22340593
  title: Aurora kinase-A inactivates DNA damage-induced apoptosis and spindle
    assembly checkpoint response functions of p73.
  findings: []
- id: PMID:22493223
  title: Structure of human Mad1 C-terminal domain reveals its involvement in
    kinetochore targeting
  findings: []
- id: PMID:24581499
  title: Nuclear pores protect genome integrity by assembling a premitotic and
    Mad1-dependent anaphase inhibitor.
  findings: []
- id: PMID:25383541
  title: The mitotic checkpoint complex binds a second CDC20 to inhibit active
    APC/C
  findings:
  - statement: MCC binds second CDC20 for APC/C inhibition
- id: PMID:25416956
  title: A proteome-scale map of the human interactome network
  findings: []
- id: PMID:25502805
  title: "A massively parallel pipeline to clone DNA variants and examine molecular phenotypes of human disease mutations."
  findings: []
- id: PMID:25852190
  title: "Integrative analysis of kinase networks in TRAIL-induced apoptosis provides a source of potential targets for combination therapy."
  findings: []
- id: PMID:26258632
  title: Mad1 promotes chromosome congression by anchoring a kinesin motor to
    the kinetochore
  findings: []
- id: PMID:26496610
  title: "A human interactome in three quantitative dimensions organized by stoichiometries and abundances."
  findings: []
- id: PMID:28514442
  title: Architecture of the human interactome defines protein communities and
    disease networks
  findings: []
- id: PMID:29997244
  title: "LuTHy: a double-readout bioluminescence-based two-hybrid technology for
    quantitative mapping of protein-protein interactions in mammalian cells."
  findings: []
- id: PMID:31467278
  title: Maximizing binary interactome mapping with a minimal number of assays
  findings: []
- id: PMID:31515488
  title: Extensive disruption of protein interactions by genetic variants across
    the allele frequency spectrum in human populations.
  findings: []
- id: PMID:32296183
  title: A reference map of the human binary protein interactome
  findings: []
- id: PMID:32814053
  title: Interactome Mapping Provides a Network of Neurodegenerative Disease
    Proteins and Uncovers Widespread Protein Aggregation in Affected Brains.
  findings: []
- id: PMID:33961781
  title: Dual proteome-scale networks reveal cell-specific remodeling of the
    human interactome
  findings: []
- id: PMID:35384245
  title: Physical and functional interactome atlas of human receptor tyrosine
    kinases
  findings: []
- id: PMID:37398436
  title: AI-guided pipeline for protein-protein interaction drug discovery
    identifies a SARS-CoV-2 inhibitor.
  findings: []
- id: PMID:37926298
  title: "Therapeutic role of 2-stearoxyphenethyl phosphocholine targeting microtubule dynamics and Wnt/β-catenin/EMT signaling in human colorectal cancer cells."
  findings: []
- id: PMID:40205054
  title: Multimodal cell maps as a foundation for structural and functional
    genomics
  findings: []
- id: PMID:9092546
  title: "Interaction of MAD2 with the carboxyl terminus of the insulin receptor but not with the IGFIR. Evidence for release from the insulin receptor after activation."
  findings:
  - statement: MAD2 interacts with insulin receptor C-terminus
- id: PMID:23509069
  title: "MISP is a novel Plk1 substrate required for proper spindle orientation and mitotic progression."
  findings: []
- id: PMID:22898774
  title: Evolution and function of the mitotic checkpoint
  findings:
  - statement: Review of SAC evolution and function
- id: PMID:19010891
  title: Role of a novel splice variant of mitotic arrest deficient 1 (MAD1),
    MAD1beta, in mitotic checkpoint control in liver cancer.
  findings:
  - statement: MAD1beta cytoplasmic localization sequesters MAD2
- id: PMID:25422469
  title: Disruption of FAT10-MAD2 binding inhibits tumor progression
  findings:
  - statement: FAT10 interaction with MAD2 affects checkpoint function
- id: PMID:18794143
  title: "HSF1 as a mitotic regulator: phosphorylation of HSF1 by Plk1 is essential
    for mitotic progression."
  findings: []
- id: PMID:21274008
  title: MTBP plays a crucial role in mitotic progression and chromosome
    segregation
  findings: []
- id: PMID:20133940
  title: Nucleoporin translocated promoter region (Tpr) associates with dynein
    complex, preventing chromosome lagging formation during mitosis.
  findings:
  - statement: TPR associates with dynein and MAD1/MAD2 during cell division
- id: PMID:19273613
  title: Spatiotemporal control of mitosis by the conserved spindle matrix
    protein Megator
  findings: []
- id: PMID:20870947
  title: Critical role of Pcid2 in B cell survival through the regulation of
    MAD2 expression.
  findings:
  - statement: Pcid2 regulates MAD2 expression in B cells
- id: PMID:19229290
  title: Dynein light intermediate chain 1 is required for progress through the
    spindle assembly checkpoint.
  findings: []
- id: PMID:10200259
  title: "A MHC-encoded ubiquitin-like protein (FAT10) binds noncovalently to the spindle assembly checkpoint protein MAD2."
  findings:
  - statement: FAT10 (MHC-encoded ubiquitin-like protein) binds MAD2
- id: PMID:15525512
  title: Phosphorylation of Cdc20 by Bub1 provides a catalytic mechanism for
    APC/C inhibition by the spindle checkpoint.
  findings: []
- id: PMID:19468067
  title: Mitotic control of kinetochore-associated dynein and spindle
    orientation by human Spindly
  findings: []
- id: PMID:17363900
  title: The human Nup107-160 nuclear pore subcomplex contributes to proper
    kinetochore functions
  findings: []
- id: Reactome:R-HSA-141409
  title: MAD1 binding to kinetochore
  findings: []
- id: Reactome:R-HSA-141422
  title: MAD2 conversion to inhibitory state
  findings: []
- id: Reactome:R-HSA-141431
  title: MAD2 association with MAD1 kinetochore complex
  findings: []
- id: Reactome:R-HSA-141439
  title: Release of activated MAD2 from kinetochores
  findings: []
- id: Reactome:R-HSA-1638803
  title: PLK1 phosphorylation of cohesin at centromeres
  findings: []
- id: Reactome:R-HSA-1638821
  title: PP2A-B56 dephosphorylation of centromeric cohesin
  findings: []
- id: Reactome:R-HSA-2467809
  title: ESPL1 (Separase) cleavage of centromeric cohesin
  findings: []
- id: Reactome:R-HSA-2467811
  title: Separation of sister chromatids
  findings: []
- id: Reactome:R-HSA-2468287
  title: CDK1 phosphorylation of CDCA5 (Sororin) at centromeres
  findings: []
- id: Reactome:R-HSA-2484822
  title: Kinetochore assembly
  findings: []
- id: Reactome:R-HSA-375302
  title: Kinetochore capture of astral microtubules
  findings: []
- id: Reactome:R-HSA-5666129
  title: CDC42:GTP recruiting DIAPH2-2 to kinetochores
  findings: []
- id: Reactome:R-HSA-5666160
  title: AURKB phosphorylation of DIAPH2-2 at kinetochores
  findings: []
- id: Reactome:R-HSA-5666169
  title: Kinetochore capture regulation by CDC42:GTP:p-DIAPH2-2
  findings: []
- id: Reactome:R-HSA-9648114
  title: EML4 recruiting NUDC to mitotic spindle
  findings: []
- id: Reactome:R-HSA-141423
  title: MCC complex binding to APC/C complex
  findings: []
- id: Reactome:R-HSA-141429
  title: APC/C inactivation via CDC20 sequestration
  findings: []
- id: Reactome:R-HSA-141437
  title: MCC complex formation
  findings: []
- id: Reactome:R-HSA-174104
  title: Ubiquitination of Cyclin A by APC/C:Cdc20
  findings: []
- id: Reactome:R-HSA-174171
  title: Cyclin A association with APC/C
  findings: []
- id: Reactome:R-HSA-174238
  title: APC/C:Cdc20 activation by MCC dissociation
  findings: []
- id: Reactome:R-HSA-174255
  title: Multiubiquitinated Cyclin A degradation
  findings: []
- id: Reactome:R-HSA-179410
  title: Nek2A association with MCC:APC/C
  findings: []
- id: Reactome:R-HSA-179417
  title: Nek2A multiubiquitination
  findings: []
- id: Reactome:R-HSA-179421
  title: Nek2A degradation
  findings: []
- id: file:human/MAD2L1/MAD2L1-deep-research-falcon.md
  title: Deep research report on MAD2L1
  findings: []

core_functions:
- molecular_function:
    id: GO:0042803
    label: protein homodimerization activity
  description: >-
    MAD2 homodimerization (specifically O-MAD2:C-MAD2 conformational heterodimer)
    is essential for the template model of MAD2 activation. Crystal structures
    demonstrate both asymmetric (O:C) and symmetric (C:C) dimers.
  directly_involved_in:
  - id: GO:0007094
    label: mitotic spindle assembly checkpoint signaling
  locations:
  - id: GO:0000776
    label: kinetochore
  in_complex:
    id: GO:1990728
    label: mitotic spindle assembly checkpoint MAD1-MAD2 complex
- molecular_function:
    id: GO:1990948
    label: ubiquitin ligase inhibitor activity
  description: >-
    MAD2 within the MCC inhibits the ubiquitin ligase activity of APC/C-CDC20,
    preventing ubiquitination of securin and cyclin B until checkpoint satisfaction.
  directly_involved_in:
  - id: GO:1904667
    label: negative regulation of ubiquitin protein ligase activity
  in_complex:
    id: GO:0033597
    label: mitotic checkpoint complex
  supported_by:
  - reference_id: PMID:10700282
    supporting_text: "The checkpoint protein Mad2 inhibits the activity of the anaphase
      promoting complex by sequestering Cdc20"
  - reference_id: PMID:11459825
    supporting_text: "We suggest that MAD2L2 and MAD2 inhibit the release of substrates
      from APC"

proposed_new_terms: []

suggested_questions:
- question: How does MAD2 conformational switching (O-MAD2 to C-MAD2) contribute
    to the amplification of checkpoint signaling?
- question: What are the structural determinants of the asymmetric O-MAD2:C-MAD2
    dimer interface?
- question: How do post-translational modifications regulate MAD2 activity and
    localization?

suggested_experiments:
- description: Live-cell imaging with MAD2 conformer-specific probes to track
    O-MAD2 to C-MAD2 conversion in real time
  hypothesis: Real-time tracking of MAD2 conformational changes will reveal the
    kinetics of checkpoint activation
- description: Reconstitution of MCC assembly on phosphorylated kinetochore
    scaffolds
  hypothesis: Phosphorylation state of kinetochore components regulates the
    efficiency of MCC assembly
- description: Structural analysis of MAD2 variants associated with checkpoint
    defects in cancer
  hypothesis: Cancer-associated MAD2 mutations disrupt key interfaces required
    for dimerization or ligand binding