dagA

Existing Annotations Review

GO Term	Evidence	Action	Reason
GO:0005886 plasma membrane	IBA GO_REF:0000033	ACCEPT	Summary: CRAC is recruited from the cytosol to the plasma membrane upon chemoattractant stimulation, where its PH domain engages PIP3. This is a well-supported and functionally central localization. Reason: Regulated translocation of CRAC to the plasma membrane (the inner leaflet, where PIP3 accumulates) is directly documented and is where CRAC exerts its adenylyl cyclase-activating function. The phylogenetic (IBA) inference agrees with direct experimental evidence. Supporting Evidence: PMID:16267269 CRAC is rapidly and transiently recruited to the plasma membrane around the entire periphery of the cell
GO:0005543 phospholipid binding	IBA GO_REF:0000033	MODIFY	Summary: CRAC binds phosphoinositides (PI(3,4,5)P3 and PI(3,4)P2) via its PH domain. The general term phospholipid binding is correct but less informative than the specific 3-phosphoinositide binding terms, which are independently annotated with direct evidence. Reason: The specific lipid ligands of CRAC are the PI3K products PIP3 and PI(3,4)P2. Replacing the generic parent with the specific PIP3-binding term better captures the molecular function established experimentally. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: GRANULARITY MISMATCH Sources checked: PANTHER:PTN008555303 · PANTHER node for PH-domain 3-phosphoinositide-binding adaptor proteins SUPPORTS SOURCE BUT NOT TARGET Generic phospholipid binding is correct but less informative than the specific PIP3 and PI(3,4)P2 binding established experimentally for CRAC UniProtKB:Q9UN19 · human DAPP1, a PIP3-binding PH-domain adaptor SUPPORTS SOURCE BUT NOT TARGET Source proteins bind 3-phosphoinositides; the specific PIP3-binding child term is preferable to the generic parent Proposed replacements: phosphatidylinositol-3,4,5-trisphosphate binding Supporting Evidence: PMID:15668169 pleckstrin homology (PH) domain-containing proteins that bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
GO:0043325 phosphatidylinositol-3,4-bisphosphate binding	IBA GO_REF:0000033	ACCEPT	Summary: The CRAC PH domain binds PI(3,4)P2 as well as PI(3,4,5)P3. This specific lipid-binding activity is directly supported. Reason: PI(3,4)P2 is one of the two PI3K products bound by the CRAC PH domain; the IBA inference matches the experimental characterization of CRAC as a 3-phosphoinositide-binding protein. Supporting Evidence: PMID:15668169 pleckstrin homology (PH) domain-containing proteins that bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
GO:0005737 cytoplasm	IEA GO_REF:0000044	ACCEPT	Summary: In unstimulated cells CRAC is a cytosolic protein, translocating to membranes only transiently upon stimulation. Cytoplasmic localization is correct. Reason: CRAC was originally purified and defined as a cytosolic regulator; its resting distribution is cytoplasmic, consistent with the UniProt subcellular location mapping. Supporting Evidence: PMID:8089184 A cytosolic protein that activates adenylyl cyclase, CRAC
GO:0046688 response to copper ion	HDA PMID:40863941 Proteomic Analysis of Heavy Metal-Induced Toxicity Using the...	MARK AS OVER ANNOTATED	Summary: This annotation derives from a 2D-gel proteomic screen in which the CRAC protein spot disappeared after copper exposure. This is a correlative change in protein abundance, not evidence that CRAC functions in a copper response. Reason: The proteomic study shows that CRAC abundance is reduced under copper stress (alongside formin-1), which is interpreted as copper disrupting aggregation-related proteins. It does not demonstrate that CRAC acts upstream of or within a physiological response to copper; the causal direction is the reverse (copper affects CRAC). Retained but flagged as over-annotation. Supporting Evidence: PMID:40863941 spot a corresponded to formin-1, spot b corresponded to CRAC PMID:40863941 Three spots (designated spots a, b, and c) were missing after Cu exposure
GO:0005829 cytosol	IDA PMID:15470246 A cell number counting factor regulates Akt/protein kinase B...	ACCEPT	Summary: Cell-fractionation and GFP-CRAC imaging assays in this study monitor the cytosol-to-membrane translocation of CRAC, consistent with a cytosolic resting pool. Reason: The study directly assays CRAC distribution between cytosol and membrane fractions, confirming the cytosolic localization of resting CRAC. Supporting Evidence: PMID:15470246 there is a quantitative change in the translocation of CRAC in the presence of anti-countin antibodies or recombinant countin
GO:0005829 cytosol	IDA PMID:15563608 Quantitative imaging of single live cells reveals spatiotemp...	ACCEPT	Summary: Live-cell imaging shows PH(Crac)-GFP returning to the cytosol after membrane translocation (adaptation), documenting the cytosolic pool. Reason: Quantitative single-cell imaging of PH(Crac)-GFP directly demonstrates cytosolic localization in the basal/adapted state. Supporting Evidence: PMID:15563608 PH domain-containing proteins evenly translocate to the plasma membrane and then quickly return to the cytosol
GO:0005829 cytosol	IDA PMID:16267269 Phosphoinositide 3-kinase activity controls the chemoattract...	ACCEPT	Summary: CRAC is defined and used here as a cytosolic (PH-domain) regulator that is recruited to membranes only on stimulation. Reason: The paper directly studies CRAC recruitment dynamics and identifies it as the cytosolic regulator of adenylyl cyclase, supporting cytosolic localization. Supporting Evidence: PMID:16267269 the pleckstrin homology (PH) domain-containing protein cytosolic regulator of adenylyl cyclase (CRAC)
GO:0005829 cytosol	IDA PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC is a cytosolic protein recruited to the membrane during chemoattractant signaling; this study established the imaging paradigm for that translocation. Reason: Directly supports the cytosolic nature of CRAC, whose recruitment from the cytosol reflects G protein activation. Supporting Evidence: PMID:9778249 transmitted by the recruitment of cytosolic proteins
GO:0005886 plasma membrane	IDA PMID:16267269 Phosphoinositide 3-kinase activity controls the chemoattract...	ACCEPT	Summary: Direct imaging shows CRAC recruited to the plasma membrane periphery upon chemoattractant stimulation. Reason: This is a core, directly observed localization where CRAC engages PIP3 and functions in ACA activation. Supporting Evidence: PMID:16267269 CRAC is rapidly and transiently recruited to the plasma membrane around the entire periphery of the cell
GO:0005886 plasma membrane	IDA PMID:23132928 Delineating the core regulatory elements crucial for directe...	ACCEPT	Summary: The PH(Crac) biosensor reports plasma-membrane phosphoinositides, confirming CRAC recruitment to the plasma membrane. Reason: PH(Crac)-GFP measures plasma-membrane PIP3/PI(3,4)P2, directly placing CRAC at the plasma membrane during chemotactic responses. Supporting Evidence: PMID:23132928 a biosensor that measures increased levels of these plasma membrane (PM) phosphoinositides
GO:0005886 plasma membrane	IDA PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC translocates to binding sites on the inner face of the plasma membrane upon G protein activation. Reason: Directly documented recruitment of CRAC to the plasma membrane (inner leaflet), a core functional localization. Supporting Evidence: PMID:9778249 The translocation of the PH domain-containing protein CRAC in D. discoideum to binding sites on the inner face of the plasma membrane
GO:0006935 chemotaxis	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: Mutational analysis shows CRAC is required for chemotaxis in addition to its role in ACA activation; a CRAC mutant unable to bind PI3K products fails to support chemotaxis. Reason: Directly demonstrated by CRAC mutants; PI3K-dependent CRAC function is required for directed migration, a core biological role. Supporting Evidence: PMID:15668169 in addition to its essential role in the activation of ACA, CRAC is involved in regulating chemotaxis PMID:15668169 A CRAC mutant that has lost the capacity to bind PI3K products does not support chemotaxis and shows minimal ACA activation
GO:0006935 chemotaxis	IMP PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC translocation to the leading edge is a hallmark of the chemotactic response; this study established leading-edge recruitment of CRAC in chemotaxing cells. Reason: The gene product's leading-edge localization during chemotaxis supports a role in directed migration. Supporting Evidence: PMID:9778249 G protein signaling events are activated at the leading edge of chemotactic cells
GO:0008047 enzyme activator activity	IMP PMID:8089184 CRAC, a cytosolic protein containing a pleckstrin homology d...	ACCEPT	Summary: CRAC is required for receptor- and G protein-mediated activation of adenylyl cyclase, acting as an adapter linking free Gbetagamma to ACA activation. This is the defining core molecular function of CRAC. Reason: The founding genetic/biochemical study shows dagA/CRAC is essential for adenylyl cyclase activation and proposes it connects Gbetagamma to the cyclase, i.e. an enzyme (adenylyl cyclase) activator role. Supporting Evidence: PMID:8089184 is required for receptor and G protein-mediated activation of adenylyl cyclase in PMID:8089184 CRAC acts to connect free G protein beta gamma subunits to adenylyl cyclase activation
GO:0009898 cytoplasmic side of plasma membrane	IDA PMID:15161938 Sensitization of Dictyostelium chemotaxis by phosphoinositid...	ACCEPT	Summary: The CRAC PH domain binds the cytoplasmic (inner) face of the plasma membrane, forming PH(Crac)-GFP patches that mark PIP3 at the leading edge. Reason: PH(Crac)-GFP localizes to inner-leaflet membrane patches, consistent with the cytoplasmic side of the plasma membrane. Supporting Evidence: PMID:15161938 translocation of PH(Crac)-GFP from the cytosol to multiple patches
GO:0016020 membrane	IDA PMID:15470246 A cell number counting factor regulates Akt/protein kinase B...	MODIFY	Summary: CRAC translocates to the membrane fraction; the general term membrane is less informative than plasma membrane, which is the actual site of CRAC recruitment. Reason: The membrane to which CRAC translocates is the plasma membrane (inner leaflet). Replacing the generic term with plasma membrane improves specificity while remaining supported by the fractionation data. Proposed replacements: plasma membrane Supporting Evidence: PMID:15470246 CRAC translocation to membranes is required for the GTPγS stimulation of adenylyl cyclase activity
GO:0016020 membrane	IDA PMID:7790361 Chemoattractant and GTP gamma S-mediated stimulation of aden...	MODIFY	Summary: Chemoattractant stimulation drives translocation of CRAC from the cytosolic to the membrane fraction. The specific compartment is the plasma membrane. Reason: The biochemical membrane fraction that CRAC translocates to corresponds to the plasma membrane, where guanine-nucleotide-generated Gbetagamma binding sites reside. Plasma membrane is the more informative term. Proposed replacements: plasma membrane Supporting Evidence: PMID:7790361 chemoattractants promote translocation of CRAC from the cytosolic to the membrane fraction
GO:0031143 pseudopodium	IDA PMID:23132928 Delineating the core regulatory elements crucial for directe...	ACCEPT	Summary: CRAC (via PH(Crac)) is among the proteins enriched in pseudopods during chemotaxis, reflecting leading-edge PIP3 accumulation. Reason: Pseudopod enrichment of PIP3-binding proteins such as CRAC is directly observed during folic-acid- and cAMP-mediated chemotaxis. Supporting Evidence: PMID:23132928 Proteins enriched in the pseudopods during chemotaxis also relocalize transiently to the plasma membrane during uniform FA stimulation
GO:0031252 cell leading edge	IDA PMID:17126332 Time-resolved responses to chemoattractant, characteristic o...	ACCEPT	Summary: CRAC accumulates at the leading edge of chemotaxing cells, marking sites of PIP3 production during front responses to chemoattractant. Reason: Leading-edge localization of CRAC/PH(Crac) is a well-established, directly imaged feature of the chemotactic response. Supporting Evidence: PMID:23132928 at the leading edge of migrating cells can be identified by the plekstrin homology (PH) domain of the cytosolic regulator of adenylyl cyclase (CRAC)
GO:0031256 leading edge membrane	IDA PMID:15563608 Quantitative imaging of single live cells reveals spatiotemp...	ACCEPT	Summary: PH(Crac)-GFP forms a highly polarized crescent at the leading-front plasma membrane in a chemoattractant gradient. Reason: Single-cell imaging directly shows CRAC translocating to the plasma membrane at the leading front, i.e. the leading edge membrane. Supporting Evidence: PMID:15563608 PH domain-containing proteins translocate from the cytosol to the plasma membrane at the leading front
GO:0031256 leading edge membrane	IDA PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC translocates to inner-face plasma membrane binding sites selectively at the stimulated (leading) edge of chemotactic cells. Reason: Directly documented leading-edge plasma-membrane recruitment of CRAC. Supporting Evidence: PMID:9778249 The translocation of the PH domain-containing protein CRAC in D. discoideum to binding sites on the inner face of the plasma membrane
GO:0106070 regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: CRAC is essential for chemoattractant-mediated activation of the aggregation adenylyl cyclase (ACA), placing it within the regulation of the adenylate cyclase-activating GPCR signaling pathway. Reason: A defining, directly demonstrated role in which CRAC is required for GPCR/PI3K-dependent activation of ACA. Supporting Evidence: PMID:15668169 CRAC is essential for the chemoattractant-mediated activation of the adenylyl cyclase ACA
GO:0106070 regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway	IMP PMID:8089184 CRAC, a cytosolic protein containing a pleckstrin homology d...	ACCEPT	Summary: The founding study shows dagA/CRAC is required for receptor and G protein-mediated activation of adenylyl cyclase. Reason: Genetic disruption of dagA abolishes receptor/G protein activation of adenylyl cyclase, a core function within this pathway. Supporting Evidence: PMID:8089184 is required for receptor and G protein-mediated activation of adenylyl cyclase in
GO:0000165 MAPK cascade	IMP PMID:9020088 The Dictyostelium mitogen-activated protein kinase ERK2 is r...	KEEP AS NON CORE	Summary: CRAC is required for proper adaptation of the MAP kinase ERK2, linking it to MAPK signaling. This is an indirect effect via cAMP/ACA signaling rather than a core CRAC function. Reason: Loss of CRAC alters ERK2 adaptation kinetics, so CRAC is functionally upstream of the ERK2 (MAPK) response. However, this is a downstream consequence of CRAC's role in cAMP signaling rather than a distinct core molecular function; retained as non-core. Supporting Evidence: PMID:9020088 CRAC, a PH domain-containing protein required for adenylyl cyclase activation, is also required for proper ERK2 adaptation
GO:0019887 protein kinase regulator activity	IMP PMID:9020088 The Dictyostelium mitogen-activated protein kinase ERK2 is r...	MARK AS OVER ANNOTATED	Summary: This molecular-function annotation is inferred from CRAC being required for ERK2 adaptation. CRAC has no known direct kinase-regulatory activity; the effect on ERK2 is indirect through cAMP signaling. Reason: CRAC is a PIP3-binding adapter, not a direct regulator of a protein kinase. The requirement for CRAC in ERK2 adaptation is mediated through its role in the cAMP/adenylyl cyclase pathway, so assigning a direct protein kinase regulator activity over-interprets the phenotype. Supporting Evidence: PMID:9020088 CRAC, a PH domain-containing protein required for adenylyl cyclase activation, is also required for proper ERK2 adaptation
GO:0001891 phagocytic cup	IDA PMID:16968738 Selective membrane exclusion in phagocytic and macropinocyti...	UNDECIDED	Summary: The cited study characterizes selective membrane sorting at phagocytic and macropinocytic cups. The cached record is abstract-only and does not mention CRAC, so the specific localization of CRAC cannot be verified here. Reason: Per policy, this experimental (IDA) curator annotation is not overruled. The cached abstract does not report CRAC, and no full text is available to confirm the CRAC-specific phagocytic-cup localization, so the annotation is left undecided pending access to the primary evidence.
GO:0070685 macropinocytic cup	IDA PMID:16968738 Selective membrane exclusion in phagocytic and macropinocyti...	UNDECIDED	Summary: As above, this study addresses membrane sorting at macropinocytic cups; the cached abstract-only record does not mention CRAC, so CRAC-specific localization cannot be verified. Reason: Cannot verify the CRAC-specific macropinocytic-cup localization from the abstract-only cached publication; the experimental annotation is retained as undecided rather than overruled.
GO:0030587 sorocarp development	HMP PMID:17659086 High-throughput analysis of spatio-temporal dynamics in Dict...	KEEP AS NON CORE	Summary: dagA-null cells are developmentally defective (developmentally null cluster). Sorocarp (fruiting body) development is disrupted because CRAC-dependent ACA activation is required for aggregation, but this is a broad downstream developmental outcome rather than CRAC's core function. Reason: High-throughput phenotyping places dagA among developmentally null mutants, so it acts within sorocarp development. This reflects the developmental consequence of losing cAMP signaling rather than a distinct molecular function; retained as non-core. Supporting Evidence: PMID:17659086 mutants in which genes such as mkpA, piaA, yakA and dagA are disrupted
GO:0007188 adenylate cyclase-modulating G protein-coupled receptor signaling pathway	IDA PMID:17606871 Locally controlled inhibitory mechanisms are involved in euk...	ACCEPT	Summary: CRAC (PH(Crac)-GFP) reports PIP3 dynamics downstream of GPCR activation and is a component of the adenylate cyclase-modulating GPCR signaling pathway. Reason: Directly observed CRAC/PIP3 responses during GPCR-mediated chemosensing support involvement in this signaling pathway. Supporting Evidence: PMID:17606871 The gradients induce a stable accumulation of the PIP3 reporter PHCrac-GFP in the front of cells near the cAMP source
GO:0043325 phosphatidylinositol-3,4-bisphosphate binding	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: CRAC binds the PI3K products PI(3,4)P2 and PI(3,4,5)P3; mutants unable to bind these lipids lose function. Reason: Directly supported lipid-binding molecular function of the CRAC PH domain; PI(3,4)P2 is one of the ligands. Supporting Evidence: PMID:15668169 pleckstrin homology (PH) domain-containing proteins that bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
GO:0048015 phosphatidylinositol-mediated signaling	TAS PMID:15366706 Chemotaxis: signalling the way forward.	KEEP AS NON CORE	Summary: CRAC is a downstream effector of PI3K-generated 3-phosphoinositides, binding them via its PH domain to transduce chemoattractant signals. It thus acts within phosphatidylinositol-mediated signaling. Reason: CRAC operates within the PI3K/PIP3 signaling branch, but the more specific and central descriptions of its role are its PIP3 binding and adenylyl cyclase activation. Retained as non-core context. Supporting Evidence: PMID:16267269 leading to the production of 3-phosphoinositides (3-PI) to which CRAC binds via its PH domain
GO:0030435 sporulation resulting in formation of a cellular spore	TAS PMID:15473840 Chemoattractant signaling in dictyostelium discoideum.	KEEP AS NON CORE	Summary: Because dagA-null cells fail to complete development, spore formation is indirectly affected. This is a distal developmental consequence of loss of CRAC-dependent aggregation signaling. Reason: dagA is among developmentally null mutants; sporulation failure follows from the block in early cAMP signaling/aggregation rather than from a direct role of CRAC in sporulation. Retained as non-core. Supporting Evidence: PMID:17659086 mutants in which genes such as mkpA, piaA, yakA and dagA are disrupted
GO:0005547 phosphatidylinositol-3,4,5-trisphosphate binding	IDA PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: The CRAC PH domain binds PI(3,4,5)P3 (PIP3); a CRAC mutant that cannot bind PI3K products loses both ACA activation and chemotaxis. This is a core molecular function. Reason: Direct evidence that PIP3 binding by CRAC is required for its function; this is the primary molecular activity of the protein. Supporting Evidence: PMID:15668169 A CRAC mutant that has lost the capacity to bind PI3K products does not support chemotaxis and shows minimal ACA activation
GO:0008064 regulation of actin polymerization or depolymerization	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	KEEP AS NON CORE	Summary: CRAC, like Akt/PKB, contributes to regulation of actin polymerization during chemotaxis downstream of PIP3. This is a secondary role relative to ACA activation. Reason: PIP3-binding effectors including CRAC influence actin dynamics during directed migration, but this is one downstream branch of CRAC signaling rather than its defining function; retained as non-core. Supporting Evidence: PMID:17606871 Both CRAC and Akt/PKB play roles in the regulation of actin polymerization during chemotaxis
GO:0031152 aggregation involved in sorocarp development	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	KEEP AS NON CORE	Summary: CRAC is essential for activation of ACA, which produces the cAMP that drives chemotactic aggregation. Loss of CRAC blocks normal aggregation. Reason: Aggregation is the developmental process that depends on CRAC-mediated cAMP relay; it is a downstream outcome of CRAC's core signaling function. Retained as non-core. Supporting Evidence: PMID:15668169 CRAC is essential for the chemoattractant-mediated activation of the adenylyl cyclase ACA, which converts ATP into cAMP, the primary chemoattractant for D. discoideum
GO:0031152 aggregation involved in sorocarp development	IMP PMID:8089184 CRAC, a cytosolic protein containing a pleckstrin homology d...	KEEP AS NON CORE	Summary: dagA is required for normal development; re-expression of the dagA cDNA restores normal development in mutant cells, consistent with a role in aggregation. Reason: Genetic evidence links dagA to aggregation-stage development via ACA activation; downstream developmental process, retained as non-core. Supporting Evidence: PMID:8089184 the cDNA restores normal development when constitutively expressed in
GO:0007188 adenylate cyclase-modulating G protein-coupled receptor signaling pathway	IDA PMID:15563608 Quantitative imaging of single live cells reveals spatiotemp...	ACCEPT	Summary: PH(Crac)-GFP translocation dynamics directly report GPCR/G protein activation and PIP3 production, situating CRAC in the adenylate cyclase-modulating GPCR signaling pathway. Reason: Quantitative imaging of CRAC translocation as a readout of GPCR signaling supports its involvement in this pathway. Supporting Evidence: PMID:15563608 monitoring the dynamics of PH(Crac)-GFP translocation in single living cells

📄 View Raw YAML

id: P35401
gene_symbol: dagA
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:44689
  label: Dictyostelium discoideum
description: dagA encodes CRAC (Cytosolic Regulator of Adenylyl Cyclase), a
  698-residue pleckstrin homology (PH) domain-containing protein of Dictyostelium
  discoideum. CRAC is a cytosolic protein at rest that, upon chemoattractant
  (cAMP) stimulation of G protein-coupled cAMP receptors, binds the PI3K lipid
  products PI(3,4,5)P3 and PI(3,4)P2 through its N-terminal PH domain and
  translocates to the inner leaflet of the plasma membrane, accumulating at the
  leading edge of chemotaxing cells. Through this regulated membrane recruitment
  CRAC is essential for receptor- and G protein (Gbetagamma)-mediated activation
  of the aggregation-stage adenylyl cyclase (ACA), which synthesizes the cAMP
  used for intercellular signal relay. CRAC-dependent PIP3 signaling also
  contributes to directed cell migration (chemotaxis) and to regulation of the
  actin cytoskeleton, and the widely used PH(Crac)-GFP reporter marks sites of
  PIP3 production during gradient sensing. Because ACA-generated cAMP drives
  aggregation, cells lacking CRAC are developmentally defective and fail to
  aggregate normally.
existing_annotations:
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: is_active_in
  review:
    summary: CRAC is recruited from the cytosol to the plasma membrane upon
      chemoattractant stimulation, where its PH domain engages PIP3. This is a
      well-supported and functionally central localization.
    action: ACCEPT
    reason: Regulated translocation of CRAC to the plasma membrane (the inner
      leaflet, where PIP3 accumulates) is directly documented and is where CRAC
      exerts its adenylyl cyclase-activating function. The phylogenetic (IBA)
      inference agrees with direct experimental evidence.
    supported_by:
    - reference_id: PMID:16267269
      supporting_text: CRAC is rapidly and transiently recruited to the plasma
        membrane around the entire periphery of the cell
- term:
    id: GO:0005543
    label: phospholipid binding
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: CRAC binds phosphoinositides (PI(3,4,5)P3 and PI(3,4)P2) via its PH
      domain. The general term phospholipid binding is correct but less
      informative than the specific 3-phosphoinositide binding terms, which are
      independently annotated with direct evidence.
    action: MODIFY
    propagation_review:
      root_cause: TERM_SCOPING_PROBLEM
      failure_modes:
      - GRANULARITY_MISMATCH
      source_entities:
      - source_id: PANTHER:PTN008555303
        source_label: "PANTHER node for PH-domain 3-phosphoinositide-binding adaptor proteins"
        source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET
        comment: "Generic phospholipid binding is correct but less informative than the specific PIP3 and PI(3,4)P2 binding established experimentally for CRAC"
      - source_id: UniProtKB:Q9UN19
        source_label: "human DAPP1, a PIP3-binding PH-domain adaptor"
        source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET
        comment: "Source proteins bind 3-phosphoinositides; the specific PIP3-binding child term is preferable to the generic parent"
    reason: The specific lipid ligands of CRAC are the PI3K products PIP3 and
      PI(3,4)P2. Replacing the generic parent with the specific PIP3-binding term
      better captures the molecular function established experimentally.
    proposed_replacement_terms:
    - id: GO:0005547
      label: phosphatidylinositol-3,4,5-trisphosphate binding
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: pleckstrin homology (PH) domain-containing proteins that
        bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
- term:
    id: GO:0043325
    label: phosphatidylinositol-3,4-bisphosphate binding
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: The CRAC PH domain binds PI(3,4)P2 as well as PI(3,4,5)P3. This
      specific lipid-binding activity is directly supported.
    action: ACCEPT
    reason: PI(3,4)P2 is one of the two PI3K products bound by the CRAC PH domain;
      the IBA inference matches the experimental characterization of CRAC as a
      3-phosphoinositide-binding protein.
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: pleckstrin homology (PH) domain-containing proteins that
        bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: In unstimulated cells CRAC is a cytosolic protein, translocating to
      membranes only transiently upon stimulation. Cytoplasmic localization is
      correct.
    action: ACCEPT
    reason: CRAC was originally purified and defined as a cytosolic regulator; its
      resting distribution is cytoplasmic, consistent with the UniProt subcellular
      location mapping.
    supported_by:
    - reference_id: PMID:8089184
      supporting_text: A cytosolic protein that activates adenylyl cyclase, CRAC
- term:
    id: GO:0046688
    label: response to copper ion
  evidence_type: HDA
  original_reference_id: PMID:40863941
  qualifier: acts_upstream_of_or_within
  review:
    summary: This annotation derives from a 2D-gel proteomic screen in which the
      CRAC protein spot disappeared after copper exposure. This is a correlative
      change in protein abundance, not evidence that CRAC functions in a copper
      response.
    action: MARK_AS_OVER_ANNOTATED
    reason: The proteomic study shows that CRAC abundance is reduced under copper
      stress (alongside formin-1), which is interpreted as copper disrupting
      aggregation-related proteins. It does not demonstrate that CRAC acts upstream
      of or within a physiological response to copper; the causal direction is the
      reverse (copper affects CRAC). Retained but flagged as over-annotation.
    supported_by:
    - reference_id: PMID:40863941
      supporting_text: spot a corresponded to formin-1, spot b corresponded to
        CRAC
    - reference_id: PMID:40863941
      supporting_text: Three spots (designated spots a, b, and c) were missing
        after Cu exposure
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: IDA
  original_reference_id: PMID:15470246
  qualifier: located_in
  review:
    summary: Cell-fractionation and GFP-CRAC imaging assays in this study monitor
      the cytosol-to-membrane translocation of CRAC, consistent with a cytosolic
      resting pool.
    action: ACCEPT
    reason: The study directly assays CRAC distribution between cytosol and
      membrane fractions, confirming the cytosolic localization of resting CRAC.
    supported_by:
    - reference_id: PMID:15470246
      supporting_text: there is a quantitative change in the translocation of CRAC
        in the presence of anti-countin antibodies or recombinant countin
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: IDA
  original_reference_id: PMID:15563608
  qualifier: located_in
  review:
    summary: Live-cell imaging shows PH(Crac)-GFP returning to the cytosol after
      membrane translocation (adaptation), documenting the cytosolic pool.
    action: ACCEPT
    reason: Quantitative single-cell imaging of PH(Crac)-GFP directly demonstrates
      cytosolic localization in the basal/adapted state.
    supported_by:
    - reference_id: PMID:15563608
      supporting_text: PH domain-containing proteins evenly translocate to the
        plasma membrane and then quickly return to the cytosol
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: IDA
  original_reference_id: PMID:16267269
  qualifier: located_in
  review:
    summary: CRAC is defined and used here as a cytosolic (PH-domain) regulator
      that is recruited to membranes only on stimulation.
    action: ACCEPT
    reason: The paper directly studies CRAC recruitment dynamics and identifies it
      as the cytosolic regulator of adenylyl cyclase, supporting cytosolic
      localization.
    supported_by:
    - reference_id: PMID:16267269
      supporting_text: the pleckstrin homology (PH) domain-containing protein
        cytosolic regulator of adenylyl cyclase (CRAC)
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: IDA
  original_reference_id: PMID:9778249
  qualifier: located_in
  review:
    summary: CRAC is a cytosolic protein recruited to the membrane during
      chemoattractant signaling; this study established the imaging paradigm for
      that translocation.
    action: ACCEPT
    reason: Directly supports the cytosolic nature of CRAC, whose recruitment from
      the cytosol reflects G protein activation.
    supported_by:
    - reference_id: PMID:9778249
      supporting_text: transmitted by the recruitment of cytosolic proteins
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IDA
  original_reference_id: PMID:16267269
  qualifier: is_active_in
  review:
    summary: Direct imaging shows CRAC recruited to the plasma membrane periphery
      upon chemoattractant stimulation.
    action: ACCEPT
    reason: This is a core, directly observed localization where CRAC engages PIP3
      and functions in ACA activation.
    supported_by:
    - reference_id: PMID:16267269
      supporting_text: CRAC is rapidly and transiently recruited to the plasma
        membrane around the entire periphery of the cell
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IDA
  original_reference_id: PMID:23132928
  qualifier: is_active_in
  review:
    summary: The PH(Crac) biosensor reports plasma-membrane phosphoinositides,
      confirming CRAC recruitment to the plasma membrane.
    action: ACCEPT
    reason: PH(Crac)-GFP measures plasma-membrane PIP3/PI(3,4)P2, directly placing
      CRAC at the plasma membrane during chemotactic responses.
    supported_by:
    - reference_id: PMID:23132928
      supporting_text: a biosensor that measures increased levels of these plasma
        membrane (PM) phosphoinositides
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IDA
  original_reference_id: PMID:9778249
  qualifier: is_active_in
  review:
    summary: CRAC translocates to binding sites on the inner face of the plasma
      membrane upon G protein activation.
    action: ACCEPT
    reason: Directly documented recruitment of CRAC to the plasma membrane
      (inner leaflet), a core functional localization.
    supported_by:
    - reference_id: PMID:9778249
      supporting_text: The translocation of the PH domain-containing protein CRAC
        in D. discoideum to binding sites on the inner face of the plasma membrane
- term:
    id: GO:0006935
    label: chemotaxis
  evidence_type: IMP
  original_reference_id: PMID:15668169
  qualifier: involved_in
  review:
    summary: Mutational analysis shows CRAC is required for chemotaxis in addition
      to its role in ACA activation; a CRAC mutant unable to bind PI3K products
      fails to support chemotaxis.
    action: ACCEPT
    reason: Directly demonstrated by CRAC mutants; PI3K-dependent CRAC function is
      required for directed migration, a core biological role.
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: in addition to its essential role in the activation of ACA,
        CRAC is involved in regulating chemotaxis
    - reference_id: PMID:15668169
      supporting_text: A CRAC mutant that has lost the capacity to bind PI3K
        products does not support chemotaxis and shows minimal ACA activation
- term:
    id: GO:0006935
    label: chemotaxis
  evidence_type: IMP
  original_reference_id: PMID:9778249
  qualifier: involved_in
  review:
    summary: CRAC translocation to the leading edge is a hallmark of the
      chemotactic response; this study established leading-edge recruitment of
      CRAC in chemotaxing cells.
    action: ACCEPT
    reason: The gene product's leading-edge localization during chemotaxis
      supports a role in directed migration.
    supported_by:
    - reference_id: PMID:9778249
      supporting_text: G protein signaling events are activated at the leading edge
        of chemotactic cells
- term:
    id: GO:0008047
    label: enzyme activator activity
  evidence_type: IMP
  original_reference_id: PMID:8089184
  qualifier: enables
  review:
    summary: CRAC is required for receptor- and G protein-mediated activation of
      adenylyl cyclase, acting as an adapter linking free Gbetagamma to ACA
      activation. This is the defining core molecular function of CRAC.
    action: ACCEPT
    reason: The founding genetic/biochemical study shows dagA/CRAC is essential for
      adenylyl cyclase activation and proposes it connects Gbetagamma to the
      cyclase, i.e. an enzyme (adenylyl cyclase) activator role.
    supported_by:
    - reference_id: PMID:8089184
      supporting_text: is required for receptor and G protein-mediated activation
        of adenylyl cyclase in
    - reference_id: PMID:8089184
      supporting_text: CRAC acts to connect free G protein beta gamma subunits to
        adenylyl cyclase activation
- term:
    id: GO:0009898
    label: cytoplasmic side of plasma membrane
  evidence_type: IDA
  original_reference_id: PMID:15161938
  qualifier: is_active_in
  review:
    summary: The CRAC PH domain binds the cytoplasmic (inner) face of the plasma
      membrane, forming PH(Crac)-GFP patches that mark PIP3 at the leading edge.
    action: ACCEPT
    reason: PH(Crac)-GFP localizes to inner-leaflet membrane patches, consistent
      with the cytoplasmic side of the plasma membrane.
    supported_by:
    - reference_id: PMID:15161938
      supporting_text: translocation of PH(Crac)-GFP from the cytosol to multiple
        patches
- term:
    id: GO:0016020
    label: membrane
  evidence_type: IDA
  original_reference_id: PMID:15470246
  qualifier: is_active_in
  review:
    summary: CRAC translocates to the membrane fraction; the general term membrane
      is less informative than plasma membrane, which is the actual site of CRAC
      recruitment.
    action: MODIFY
    reason: The membrane to which CRAC translocates is the plasma membrane (inner
      leaflet). Replacing the generic term with plasma membrane improves
      specificity while remaining supported by the fractionation data.
    proposed_replacement_terms:
    - id: GO:0005886
      label: plasma membrane
    supported_by:
    - reference_id: PMID:15470246
      supporting_text: CRAC translocation to membranes is required for the GTPγS
        stimulation of adenylyl cyclase activity
- term:
    id: GO:0016020
    label: membrane
  evidence_type: IDA
  original_reference_id: PMID:7790361
  qualifier: is_active_in
  review:
    summary: Chemoattractant stimulation drives translocation of CRAC from the
      cytosolic to the membrane fraction. The specific compartment is the plasma
      membrane.
    action: MODIFY
    reason: The biochemical membrane fraction that CRAC translocates to corresponds
      to the plasma membrane, where guanine-nucleotide-generated Gbetagamma binding
      sites reside. Plasma membrane is the more informative term.
    proposed_replacement_terms:
    - id: GO:0005886
      label: plasma membrane
    supported_by:
    - reference_id: PMID:7790361
      supporting_text: chemoattractants promote translocation of CRAC from the
        cytosolic to the membrane fraction
- term:
    id: GO:0031143
    label: pseudopodium
  evidence_type: IDA
  original_reference_id: PMID:23132928
  qualifier: is_active_in
  review:
    summary: CRAC (via PH(Crac)) is among the proteins enriched in pseudopods
      during chemotaxis, reflecting leading-edge PIP3 accumulation.
    action: ACCEPT
    reason: Pseudopod enrichment of PIP3-binding proteins such as CRAC is directly
      observed during folic-acid- and cAMP-mediated chemotaxis.
    supported_by:
    - reference_id: PMID:23132928
      supporting_text: Proteins enriched in the pseudopods during chemotaxis also
        relocalize transiently to the plasma membrane during uniform FA stimulation
- term:
    id: GO:0031252
    label: cell leading edge
  evidence_type: IDA
  original_reference_id: PMID:17126332
  qualifier: is_active_in
  review:
    summary: CRAC accumulates at the leading edge of chemotaxing cells, marking
      sites of PIP3 production during front responses to chemoattractant.
    action: ACCEPT
    reason: Leading-edge localization of CRAC/PH(Crac) is a well-established,
      directly imaged feature of the chemotactic response.
    supported_by:
    - reference_id: PMID:23132928
      supporting_text: at the leading edge of migrating cells can be identified by
        the plekstrin homology (PH) domain of the cytosolic regulator of adenylyl
        cyclase (CRAC)
- term:
    id: GO:0031256
    label: leading edge membrane
  evidence_type: IDA
  original_reference_id: PMID:15563608
  qualifier: is_active_in
  review:
    summary: PH(Crac)-GFP forms a highly polarized crescent at the leading-front
      plasma membrane in a chemoattractant gradient.
    action: ACCEPT
    reason: Single-cell imaging directly shows CRAC translocating to the plasma
      membrane at the leading front, i.e. the leading edge membrane.
    supported_by:
    - reference_id: PMID:15563608
      supporting_text: PH domain-containing proteins translocate from the cytosol
        to the plasma membrane at the leading front
- term:
    id: GO:0031256
    label: leading edge membrane
  evidence_type: IDA
  original_reference_id: PMID:9778249
  qualifier: is_active_in
  review:
    summary: CRAC translocates to inner-face plasma membrane binding sites
      selectively at the stimulated (leading) edge of chemotactic cells.
    action: ACCEPT
    reason: Directly documented leading-edge plasma-membrane recruitment of CRAC.
    supported_by:
    - reference_id: PMID:9778249
      supporting_text: The translocation of the PH domain-containing protein CRAC
        in D. discoideum to binding sites on the inner face of the plasma membrane
- term:
    id: GO:0106070
    label: regulation of adenylate cyclase-activating G protein-coupled receptor
      signaling pathway
  evidence_type: IMP
  original_reference_id: PMID:15668169
  qualifier: involved_in
  review:
    summary: CRAC is essential for chemoattractant-mediated activation of the
      aggregation adenylyl cyclase (ACA), placing it within the regulation of the
      adenylate cyclase-activating GPCR signaling pathway.
    action: ACCEPT
    reason: A defining, directly demonstrated role in which CRAC is required for
      GPCR/PI3K-dependent activation of ACA.
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: CRAC is essential for the chemoattractant-mediated
        activation of the adenylyl cyclase ACA
- term:
    id: GO:0106070
    label: regulation of adenylate cyclase-activating G protein-coupled receptor
      signaling pathway
  evidence_type: IMP
  original_reference_id: PMID:8089184
  qualifier: involved_in
  review:
    summary: The founding study shows dagA/CRAC is required for receptor and G
      protein-mediated activation of adenylyl cyclase.
    action: ACCEPT
    reason: Genetic disruption of dagA abolishes receptor/G protein activation of
      adenylyl cyclase, a core function within this pathway.
    supported_by:
    - reference_id: PMID:8089184
      supporting_text: is required for receptor and G protein-mediated activation
        of adenylyl cyclase in
- term:
    id: GO:0000165
    label: MAPK cascade
  evidence_type: IMP
  original_reference_id: PMID:9020088
  qualifier: involved_in
  review:
    summary: CRAC is required for proper adaptation of the MAP kinase ERK2, linking
      it to MAPK signaling. This is an indirect effect via cAMP/ACA signaling
      rather than a core CRAC function.
    action: KEEP_AS_NON_CORE
    reason: Loss of CRAC alters ERK2 adaptation kinetics, so CRAC is functionally
      upstream of the ERK2 (MAPK) response. However, this is a downstream
      consequence of CRAC's role in cAMP signaling rather than a distinct core
      molecular function; retained as non-core.
    supported_by:
    - reference_id: PMID:9020088
      supporting_text: CRAC, a PH domain-containing protein required for adenylyl
        cyclase activation, is also required for proper ERK2 adaptation
- term:
    id: GO:0019887
    label: protein kinase regulator activity
  evidence_type: IMP
  original_reference_id: PMID:9020088
  qualifier: enables
  review:
    summary: This molecular-function annotation is inferred from CRAC being
      required for ERK2 adaptation. CRAC has no known direct kinase-regulatory
      activity; the effect on ERK2 is indirect through cAMP signaling.
    action: MARK_AS_OVER_ANNOTATED
    reason: CRAC is a PIP3-binding adapter, not a direct regulator of a protein
      kinase. The requirement for CRAC in ERK2 adaptation is mediated through its
      role in the cAMP/adenylyl cyclase pathway, so assigning a direct protein
      kinase regulator activity over-interprets the phenotype.
    supported_by:
    - reference_id: PMID:9020088
      supporting_text: CRAC, a PH domain-containing protein required for adenylyl
        cyclase activation, is also required for proper ERK2 adaptation
- term:
    id: GO:0001891
    label: phagocytic cup
  evidence_type: IDA
  original_reference_id: PMID:16968738
  qualifier: located_in
  review:
    summary: The cited study characterizes selective membrane sorting at phagocytic
      and macropinocytic cups. The cached record is abstract-only and does not
      mention CRAC, so the specific localization of CRAC cannot be verified here.
    action: UNDECIDED
    reason: Per policy, this experimental (IDA) curator annotation is not
      overruled. The cached abstract does not report CRAC, and no full text is
      available to confirm the CRAC-specific phagocytic-cup localization, so the
      annotation is left undecided pending access to the primary evidence.
- term:
    id: GO:0070685
    label: macropinocytic cup
  evidence_type: IDA
  original_reference_id: PMID:16968738
  qualifier: located_in
  review:
    summary: As above, this study addresses membrane sorting at macropinocytic
      cups; the cached abstract-only record does not mention CRAC, so
      CRAC-specific localization cannot be verified.
    action: UNDECIDED
    reason: Cannot verify the CRAC-specific macropinocytic-cup localization from
      the abstract-only cached publication; the experimental annotation is retained
      as undecided rather than overruled.
- term:
    id: GO:0030587
    label: sorocarp development
  evidence_type: HMP
  original_reference_id: PMID:17659086
  qualifier: acts_upstream_of_or_within
  review:
    summary: dagA-null cells are developmentally defective (developmentally null
      cluster). Sorocarp (fruiting body) development is disrupted because
      CRAC-dependent ACA activation is required for aggregation, but this is a
      broad downstream developmental outcome rather than CRAC's core function.
    action: KEEP_AS_NON_CORE
    reason: High-throughput phenotyping places dagA among developmentally null
      mutants, so it acts within sorocarp development. This reflects the
      developmental consequence of losing cAMP signaling rather than a distinct
      molecular function; retained as non-core.
    supported_by:
    - reference_id: PMID:17659086
      supporting_text: mutants in which genes such as mkpA, piaA, yakA and dagA are
        disrupted
- term:
    id: GO:0007188
    label: adenylate cyclase-modulating G protein-coupled receptor signaling pathway
  evidence_type: IDA
  original_reference_id: PMID:17606871
  qualifier: involved_in
  review:
    summary: CRAC (PH(Crac)-GFP) reports PIP3 dynamics downstream of GPCR
      activation and is a component of the adenylate cyclase-modulating GPCR
      signaling pathway.
    action: ACCEPT
    reason: Directly observed CRAC/PIP3 responses during GPCR-mediated chemosensing
      support involvement in this signaling pathway.
    supported_by:
    - reference_id: PMID:17606871
      supporting_text: The gradients induce a stable accumulation of the PIP3
        reporter PHCrac-GFP in the front of cells near the cAMP source
- term:
    id: GO:0043325
    label: phosphatidylinositol-3,4-bisphosphate binding
  evidence_type: IMP
  original_reference_id: PMID:15668169
  qualifier: enables
  review:
    summary: CRAC binds the PI3K products PI(3,4)P2 and PI(3,4,5)P3; mutants unable
      to bind these lipids lose function.
    action: ACCEPT
    reason: Directly supported lipid-binding molecular function of the CRAC PH
      domain; PI(3,4)P2 is one of the ligands.
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: pleckstrin homology (PH) domain-containing proteins that
        bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
- term:
    id: GO:0048015
    label: phosphatidylinositol-mediated signaling
  evidence_type: TAS
  original_reference_id: PMID:15366706
  qualifier: acts_upstream_of_or_within
  review:
    summary: CRAC is a downstream effector of PI3K-generated 3-phosphoinositides,
      binding them via its PH domain to transduce chemoattractant signals. It thus
      acts within phosphatidylinositol-mediated signaling.
    action: KEEP_AS_NON_CORE
    reason: CRAC operates within the PI3K/PIP3 signaling branch, but the more
      specific and central descriptions of its role are its PIP3 binding and
      adenylyl cyclase activation. Retained as non-core context.
    supported_by:
    - reference_id: PMID:16267269
      supporting_text: leading to the production of 3-phosphoinositides (3-PI) to
        which CRAC binds via its PH domain
- term:
    id: GO:0030435
    label: sporulation resulting in formation of a cellular spore
  evidence_type: TAS
  original_reference_id: PMID:15473840
  qualifier: acts_upstream_of_or_within
  review:
    summary: Because dagA-null cells fail to complete development, spore formation
      is indirectly affected. This is a distal developmental consequence of loss of
      CRAC-dependent aggregation signaling.
    action: KEEP_AS_NON_CORE
    reason: dagA is among developmentally null mutants; sporulation failure follows
      from the block in early cAMP signaling/aggregation rather than from a direct
      role of CRAC in sporulation. Retained as non-core.
    supported_by:
    - reference_id: PMID:17659086
      supporting_text: mutants in which genes such as mkpA, piaA, yakA and dagA are
        disrupted
- term:
    id: GO:0005547
    label: phosphatidylinositol-3,4,5-trisphosphate binding
  evidence_type: IDA
  original_reference_id: PMID:15668169
  qualifier: enables
  review:
    summary: The CRAC PH domain binds PI(3,4,5)P3 (PIP3); a CRAC mutant that cannot
      bind PI3K products loses both ACA activation and chemotaxis. This is a core
      molecular function.
    action: ACCEPT
    reason: Direct evidence that PIP3 binding by CRAC is required for its function;
      this is the primary molecular activity of the protein.
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: A CRAC mutant that has lost the capacity to bind PI3K
        products does not support chemotaxis and shows minimal ACA activation
- term:
    id: GO:0008064
    label: regulation of actin polymerization or depolymerization
  evidence_type: IMP
  original_reference_id: PMID:15668169
  qualifier: acts_upstream_of_or_within
  review:
    summary: CRAC, like Akt/PKB, contributes to regulation of actin polymerization
      during chemotaxis downstream of PIP3. This is a secondary role relative to
      ACA activation.
    action: KEEP_AS_NON_CORE
    reason: PIP3-binding effectors including CRAC influence actin dynamics during
      directed migration, but this is one downstream branch of CRAC signaling
      rather than its defining function; retained as non-core.
    supported_by:
    - reference_id: PMID:17606871
      supporting_text: Both CRAC and Akt/PKB play roles in the regulation of actin
        polymerization during chemotaxis
- term:
    id: GO:0031152
    label: aggregation involved in sorocarp development
  evidence_type: IMP
  original_reference_id: PMID:15668169
  qualifier: acts_upstream_of_or_within
  review:
    summary: CRAC is essential for activation of ACA, which produces the cAMP that
      drives chemotactic aggregation. Loss of CRAC blocks normal aggregation.
    action: KEEP_AS_NON_CORE
    reason: Aggregation is the developmental process that depends on CRAC-mediated
      cAMP relay; it is a downstream outcome of CRAC's core signaling function.
      Retained as non-core.
    supported_by:
    - reference_id: PMID:15668169
      supporting_text: CRAC is essential for the chemoattractant-mediated
        activation of the adenylyl cyclase ACA, which converts ATP into cAMP, the
        primary chemoattractant for D. discoideum
- term:
    id: GO:0031152
    label: aggregation involved in sorocarp development
  evidence_type: IMP
  original_reference_id: PMID:8089184
  qualifier: acts_upstream_of_or_within
  review:
    summary: dagA is required for normal development; re-expression of the dagA cDNA
      restores normal development in mutant cells, consistent with a role in
      aggregation.
    action: KEEP_AS_NON_CORE
    reason: Genetic evidence links dagA to aggregation-stage development via ACA
      activation; downstream developmental process, retained as non-core.
    supported_by:
    - reference_id: PMID:8089184
      supporting_text: the cDNA restores normal development when constitutively
        expressed in
- term:
    id: GO:0007188
    label: adenylate cyclase-modulating G protein-coupled receptor signaling pathway
  evidence_type: IDA
  original_reference_id: PMID:15563608
  qualifier: involved_in
  review:
    summary: PH(Crac)-GFP translocation dynamics directly report GPCR/G protein
      activation and PIP3 production, situating CRAC in the adenylate
      cyclase-modulating GPCR signaling pathway.
    action: ACCEPT
    reason: Quantitative imaging of CRAC translocation as a readout of GPCR
      signaling supports its involvement in this pathway.
    supported_by:
    - reference_id: PMID:15563608
      supporting_text: monitoring the dynamics of PH(Crac)-GFP translocation in
        single living cells
references:
- id: GO_REF:0000033
  title: Annotation inferences using phylogenetic trees
  findings: []
- id: GO_REF:0000044
  title: Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location
    vocabulary mapping, accompanied by conservative changes to GO terms applied by
    UniProt
  findings: []
- id: PMID:15161938
  title: Sensitization of Dictyostelium chemotaxis by phosphoinositide-3-kinase-mediated
    self-organizing signalling patches.
  findings:
  - statement: PH(Crac)-GFP marks the leading edge of Dictyostelium cells in
      chemoattractant gradients and forms self-organizing membrane patches.
    supporting_text: translocation of PH(Crac)-GFP from the cytosol to multiple
      patches
- id: PMID:15366706
  title: 'Chemotaxis: signalling the way forward.'
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Review of chemotaxis signaling; supports general context but does
      not itself provide CRAC-specific molecular evidence.
- id: PMID:15470246
  title: A cell number counting factor regulates Akt/protein kinase B to regulate
    Dictyostelium discoideum group size.
  findings:
  - statement: CRAC translocation between cytosol and membranes was directly assayed
      by fractionation and GFP-CRAC imaging.
    supporting_text: there is a quantitative change in the translocation of CRAC in
      the presence of anti-countin antibodies or recombinant countin
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Primarily an Akt/PKB study, but the full text directly assays CRAC
      translocation, supporting cytosol/membrane localization annotations.
- id: PMID:15473840
  title: Chemoattractant signaling in dictyostelium discoideum.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Review placing CRAC within chemoattractant signaling modules;
      supports developmental/sporulation context indirectly.
- id: PMID:15563608
  title: Quantitative imaging of single live cells reveals spatiotemporal dynamics
    of multistep signaling events of chemoattractant gradient sensing in Dictyostelium.
  findings:
  - statement: PH(Crac)-GFP translocation from cytosol to the leading-front plasma
      membrane reports PIP3 dynamics during gradient sensing.
    supporting_text: PH domain-containing proteins translocate from the cytosol to
      the plasma membrane at the leading front
- id: PMID:15668169
  title: The PI3K-mediated activation of CRAC independently regulates adenylyl cyclase
    activation and chemotaxis.
  findings:
  - statement: CRAC is essential for chemoattractant-mediated activation of adenylyl
      cyclase ACA and is separately required for chemotaxis.
    supporting_text: in addition to its essential role in the activation of ACA,
      CRAC is involved in regulating chemotaxis
  - statement: A CRAC mutant unable to bind PI3K products loses chemotaxis and ACA
      activation, showing PIP3 binding is required for CRAC function.
    supporting_text: A CRAC mutant that has lost the capacity to bind PI3K products
      does not support chemotaxis and shows minimal ACA activation
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Key mutational study separating the ACA-activation and chemotaxis
      functions of CRAC downstream of PI3K.
- id: PMID:16267269
  title: Phosphoinositide 3-kinase activity controls the chemoattractant-mediated
    activation and adaptation of adenylyl cyclase.
  findings:
  - statement: CRAC is rapidly recruited to the plasma membrane on chemoattractant
      stimulation and binds 3-phosphoinositides via its PH domain.
    supporting_text: CRAC is rapidly and transiently recruited to the plasma
      membrane around the entire periphery of the cell
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Full text directly describes CRAC recruitment dynamics and its
      3-phosphoinositide binding.
- id: PMID:16968738
  title: Selective membrane exclusion in phagocytic and macropinocytic cups.
  findings: []
  reference_review:
    relevance: LOW
    correctness: UNVERIFIED
    review_notes: Cached record is abstract-only and does not mention CRAC; cannot
      verify the CRAC-specific phagocytic/macropinocytic-cup localizations.
- id: PMID:17126332
  title: Time-resolved responses to chemoattractant, characteristic of the front and
    tail of Dictyostelium cells.
  findings: []
- id: PMID:17606871
  title: Locally controlled inhibitory mechanisms are involved in eukaryotic GPCR-mediated
    chemosensing.
  findings:
  - statement: The PIP3 reporter PHCrac-GFP accumulates at the front of cells in a
      cAMP gradient, and CRAC contributes to actin regulation during chemotaxis.
    supporting_text: Both CRAC and Akt/PKB play roles in the regulation of actin
      polymerization during chemotaxis
- id: PMID:17659086
  title: High-throughput analysis of spatio-temporal dynamics in Dictyostelium.
  findings:
  - statement: dagA is among the developmentally null mutants identified by
      high-throughput phenotyping.
    supporting_text: mutants in which genes such as mkpA, piaA, yakA and dagA are
      disrupted
- id: PMID:23132928
  title: Delineating the core regulatory elements crucial for directed cell migration
    by examining folic-acid-mediated responses.
  findings:
  - statement: The PH domain of CRAC is a biosensor for leading-edge plasma-membrane
      phosphoinositides, and pseudopod-enriched proteins relocalize to the plasma
      membrane on uniform stimulation.
    supporting_text: a biosensor that measures increased levels of these plasma
      membrane (PM) phosphoinositides
- id: PMID:40863941
  title: 'Proteomic Analysis of Heavy Metal-Induced Toxicity Using the Cellular Slime
    Mould Dictyostelium discoideum: Effects of Copper Exposure on Aggregation and
    Protein Expression.'
  findings:
  - statement: The CRAC protein spot disappeared from 2D gels after copper exposure,
      correlating copper toxicity with reduced CRAC abundance.
    supporting_text: spot a corresponded to formin-1, spot b corresponded to CRAC
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Correlative proteomics; supports a change in CRAC abundance under
      copper stress but not a functional role in copper response.
- id: PMID:7790361
  title: Chemoattractant and GTP gamma S-mediated stimulation of adenylyl cyclase
    in Dictyostelium requires translocation of CRAC to membranes.
  findings:
  - statement: Chemoattractant stimulation drives translocation of CRAC from the
      cytosolic to the membrane fraction, required for adenylyl cyclase activation.
    supporting_text: chemoattractants promote translocation of CRAC from the
      cytosolic to the membrane fraction
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Establishes that CRAC membrane translocation is required for
      chemoattractant/GTPgammaS activation of adenylyl cyclase.
- id: PMID:8089184
  title: CRAC, a cytosolic protein containing a pleckstrin homology domain, is required
    for receptor and G protein-mediated activation of adenylyl cyclase in Dictyostelium.
  findings:
  - statement: dagA is the structural gene for CRAC and is required for receptor and
      G protein-mediated activation of adenylyl cyclase.
    supporting_text: is required for receptor and G protein-mediated activation of
      adenylyl cyclase in
  - statement: CRAC is proposed to connect free Gbetagamma subunits to adenylyl
      cyclase activation.
    supporting_text: CRAC acts to connect free G protein beta gamma subunits to
      adenylyl cyclase activation
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Founding genetic study identifying dagA/CRAC and its essential
      role in adenylyl cyclase activation.
- id: PMID:9020088
  title: The Dictyostelium mitogen-activated protein kinase ERK2 is regulated by Ras
    and cAMP-dependent protein kinase (PKA) and mediates PKA function.
  findings:
  - statement: CRAC is required for proper adaptation of the MAP kinase ERK2.
    supporting_text: CRAC, a PH domain-containing protein required for adenylyl
      cyclase activation, is also required for proper ERK2 adaptation
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Supports an indirect link between CRAC and ERK2 adaptation; does
      not establish direct protein kinase regulator activity.
- id: PMID:9778249
  title: G protein signaling events are activated at the leading edge of chemotactic
    cells.
  findings:
  - statement: CRAC translocates to inner-face plasma membrane binding sites
      selectively at the leading edge in a chemoattractant gradient.
    supporting_text: The translocation of the PH domain-containing protein CRAC in
      D. discoideum to binding sites on the inner face of the plasma membrane
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Established the CRAC/PH(Crac) imaging paradigm for leading-edge G
      protein signaling during chemotaxis.
core_functions:
- description: CRAC is a cytosolic PH-domain adapter that, upon chemoattractant
    stimulation, binds the PI3K products PI(3,4,5)P3 and PI(3,4)P2 and translocates
    to the plasma membrane (leading-edge inner leaflet), coupling GPCR/PI3K
    signaling to downstream effectors.
  molecular_function:
    id: GO:0005547
    label: phosphatidylinositol-3,4,5-trisphosphate binding
  locations:
  - id: GO:0005829
    label: cytosol
  - id: GO:0005886
    label: plasma membrane
  - id: GO:0031256
    label: leading edge membrane
  directly_involved_in:
  - id: GO:0007188
    label: adenylate cyclase-modulating G protein-coupled receptor signaling pathway
  - id: GO:0006935
    label: chemotaxis
  supported_by:
  - reference_id: PMID:15668169
    supporting_text: A CRAC mutant that has lost the capacity to bind PI3K products
      does not support chemotaxis and shows minimal ACA activation
  - reference_id: PMID:16267269
    supporting_text: CRAC is rapidly and transiently recruited to the plasma
      membrane around the entire periphery of the cell
- description: CRAC is required for receptor- and G protein (Gbetagamma)-mediated
    activation of the aggregation-stage adenylyl cyclase (ACA), functioning as an
    activator that links activated G protein signaling to cAMP synthesis for
    intercellular signal relay.
  molecular_function:
    id: GO:0008047
    label: enzyme activator activity
  locations:
  - id: GO:0005886
    label: plasma membrane
  directly_involved_in:
  - id: GO:0106070
    label: regulation of adenylate cyclase-activating G protein-coupled receptor
      signaling pathway
  supported_by:
  - reference_id: PMID:8089184
    supporting_text: CRAC acts to connect free G protein beta gamma subunits to
      adenylyl cyclase activation
  - reference_id: PMID:15668169
    supporting_text: CRAC is essential for the chemoattractant-mediated activation
      of the adenylyl cyclase ACA

Gene Description

Existing Annotations Review

Core Functions

CRAC is a cytosolic PH-domain adapter that, upon chemoattractant stimulation, binds the PI3K products PI(3,4,5)P3 and PI(3,4)P2 and translocates to the plasma membrane (leading-edge inner leaflet), coupling GPCR/PI3K signaling to downstream effectors.

CRAC is required for receptor- and G protein (Gbetagamma)-mediated activation of the aggregation-stage adenylyl cyclase (ACA), functioning as an activator that links activated G protein signaling to cAMP synthesis for intercellular signal relay.

References

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