id: P33316
gene_symbol: DUT
product_type: PROTEIN
status: INITIALIZED
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: >-
  DUT encodes deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase; dUTP
  pyrophosphatase; EC 3.6.1.23), a Mg2+-dependent hydrolase that cleaves dUTP into
  dUMP and inorganic diphosphate. Through this single reaction the enzyme performs a
  dual, complementary role in pyrimidine deoxyribonucleotide metabolism: it keeps the
  intracellular dUTP:dTTP ratio low, preventing uracil misincorporation into DNA (and
  the resulting futile base-excision-repair cycles and DNA strand breaks), and it
  produces dUMP, the substrate that thymidylate synthase methylates to dTMP during de
  novo thymidylate biosynthesis. The active enzyme is a homotrimer whose substrate
  sites are formed at subunit interfaces, with catalytic residues contributed by all
  three subunits. In humans, alternative splicing of the N-terminus yields a nuclear
  isoform (DUT-N) and a mitochondrion-targeted isoform (DUT-M) bearing a cleavable
  transit peptide, so that dUTPase activity is provided in both the nucleus/cytosol
  and the mitochondrion. The nuclear isoform is cell-cycle regulated, peaking with
  DNA replication. dUTPase is essential for genome integrity; biallelic
  loss-of-function variants cause a bone marrow failure and diabetes mellitus syndrome.
alternative_products:
- name: 3 (DUT-M)
  id: P33316-3
- name: 2 (DUT-N)
  id: P33316-2
  sequence_note: VSP_001324
existing_annotations:
- term:
    id: GO:0004170
    label: dUTP diphosphatase activity
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: >-
      Phylogenetically inferred dUTP diphosphatase (dUTPase) activity. This is the
      well-established, experimentally confirmed core molecular function of DUT and is
      supported at multiple levels of evidence.
    action: ACCEPT
    reason: >-
      dUTP diphosphatase activity is the defining, experimentally proven function of
      human dUTPase, confirmed by kinetics and crystal structures of the substrate/Mg2+
      complex. The IBA is consistent with the direct experimental annotations and with
      the dUTPase family assignment.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        dUTPase hydrolyzes dUTP to dUMP and pyrophosphate, simultaneously reducing
        dUTP levels and providing the dUMP for dTTP biosynthesis.
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: "Belongs to the dUTPase family"
- term:
    id: GO:0000287
    label: magnesium ion binding
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: >-
      Phylogenetically inferred magnesium ion binding. dUTPase is a Mg2+-dependent
      enzyme; the catalytic reaction requires a bound Mg2+ ion, confirmed in the
      crystal structures.
    action: ACCEPT
    reason: >-
      Mg2+ is an experimentally confirmed cofactor of human dUTPase (UniProt COFACTOR,
      from PubMed:8805593), and structures were solved as the enzyme:substrate:Mg
      complex. The IBA is well supported.
    supported_by:
    - reference_id: PMID:17880943
      supporting_text: >-
        We determined the crystal structure of the
        enzyme:alpha,beta-imino-dUTP:Mg complex
- term:
    id: GO:0006226
    label: dUMP biosynthetic process
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: involved_in
  review:
    summary: >-
      Phylogenetically inferred involvement in dUMP biosynthesis. Hydrolysis of dUTP
      by dUTPase is the direct source of dUMP, the immediate precursor of dTMP.
    action: ACCEPT
    reason: >-
      dUMP is the direct product of the dUTPase reaction and the substrate for
      thymidylate synthase; this is a core biological role of DUT and is captured by
      UniPathway "dUMP biosynthesis; dUMP from dCTP (dUTP route)".
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        dUTPase hydrolyzes dUTP to dUMP and pyrophosphate, simultaneously reducing
        dUTP levels and providing the dUMP for dTTP biosynthesis.
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: "Pyrimidine metabolism; dUMP biosynthesis; dUMP from dCTP"
- term:
    id: GO:0046081
    label: dUTP catabolic process
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: involved_in
  review:
    summary: >-
      Phylogenetically inferred involvement in dUTP catabolism. The dUTPase reaction
      is the catabolic step that removes dUTP from the deoxyribonucleotide pool.
    action: ACCEPT
    reason: >-
      dUTP catabolism (hydrolysis of dUTP) is exactly what dUTPase does; this is a
      core biological-process framing of the enzymatic activity and is well supported.
    supported_by:
    - reference_id: PMID:15322254
      supporting_text: >-
        dUTPase, which eliminates dUTP from the DNA biosynthetic pathway, opposes
        uracil misincorporation
- term:
    id: GO:0000287
    label: magnesium ion binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000002
  qualifier: enables
  review:
    summary: >-
      InterPro2GO electronic transfer of magnesium ion binding from the dUTPase
      InterPro signature (IPR008181). Consistent with the experimentally confirmed
      Mg2+ cofactor.
    action: ACCEPT
    reason: >-
      The InterPro-based IEA reproduces the experimentally validated Mg2+ dependence
      of dUTPase and is redundant with, and consistent with, the IBA magnesium ion
      binding annotation.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        The dUTPase nucleotide-binding sites incorporate some features of other
        nucleotide-binding proteins
- term:
    id: GO:0004170
    label: dUTP diphosphatase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: >-
      Combined-method electronic annotation of dUTP diphosphatase activity (ARBA /
      EC:3.6.1.23 / RHEA:10248 / InterPro). Matches the experimentally proven function.
    action: ACCEPT
    reason: >-
      This IEA reproduces the core, experimentally confirmed enzymatic activity of DUT
      and is fully consistent with the EXP/IDA/IBA annotations for the same term.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        dUTPase hydrolyzes dUTP to dUMP and pyrophosphate
- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: >-
      UniProt Subcellular-Location keyword mapping to nucleus, corresponding to the
      nuclear isoform (DUT-N / isoform 2).
    action: ACCEPT
    reason: >-
      Nuclear localization of the nuclear isoform is experimentally established
      (PubMed:8631816), so this SubCell-derived IEA is correct.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the lower molecular weight form of dUTPase (DUT-N) is associated with the
        nucleus
- term:
    id: GO:0005739
    label: mitochondrion
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: >-
      UniProt Subcellular-Location keyword mapping to mitochondrion, corresponding to
      the mitochondrial isoform (DUT-M / isoform 3) with its N-terminal transit peptide.
    action: ACCEPT
    reason: >-
      Mitochondrial localization of the DUT-M isoform is experimentally established
      (PubMed:8631816) and independently supported by high-confidence mitochondrial
      proteomics; this SubCell-derived IEA is correct.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the higher molecular weight species (DUT-M) fractionates with the mitochondria
- term:
    id: GO:0006226
    label: dUMP biosynthetic process
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: involved_in
  review:
    summary: >-
      Combined-method electronic annotation to dUMP biosynthetic process, mirroring
      the IBA involvement. dUMP is the direct product of dUTPase.
    action: ACCEPT
    reason: >-
      Correct and redundant with the IBA dUMP biosynthesis annotation; dUMP is the
      immediate product of the dUTPase reaction.
    supported_by:
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: "Pyrimidine metabolism; dUMP biosynthesis; dUMP from dCTP"
- term:
    id: GO:0009177
    label: pyrimidine deoxyribonucleoside monophosphate biosynthetic process
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  qualifier: involved_in
  review:
    summary: >-
      ARBA electronic annotation to a broader parent process (pyrimidine
      deoxyribonucleoside monophosphate biosynthesis). This generalizes the specific
      dUMP biosynthetic process role.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Not wrong (dUMP is a pyrimidine deoxyribonucleoside monophosphate), but this ARBA
      IEA is less precise than the dedicated GO:0006226 dUMP biosynthetic process term
      already annotated; the specific term better captures DUT's role.
    supported_by:
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: "Pyrimidine metabolism; dUMP biosynthesis; dUMP from dCTP"
- term:
    id: GO:0009221
    label: pyrimidine deoxyribonucleotide biosynthetic process
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  qualifier: involved_in
  review:
    summary: >-
      ARBA electronic annotation to a broad pyrimidine deoxyribonucleotide biosynthetic
      process. dUTPase supplies dUMP that feeds dTMP/dTTP biosynthesis.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Directionally correct as DUT feeds pyrimidine deoxyribonucleotide (dTMP/dTTP)
      biosynthesis by providing dUMP, but this is a broad parent term less precise than
      the specific dUMP biosynthetic process / dTMP biosynthetic process annotations
      already present.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        providing the dUMP for dTTP biosynthesis
- term:
    id: GO:0046081
    label: dUTP catabolic process
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: involved_in
  review:
    summary: >-
      Combined-method electronic annotation to dUTP catabolic process, mirroring the
      IBA involvement.
    action: ACCEPT
    reason: >-
      Correct and redundant with the IBA dUTP catabolic process annotation; hydrolysis
      of dUTP is the catabolic reaction catalyzed by dUTPase.
    supported_by:
    - reference_id: PMID:15322254
      supporting_text: >-
        dUTPase, which eliminates dUTP from the DNA biosynthetic pathway
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:16189514
  qualifier: enables
  review:
    summary: >-
      IntAct-curated binary interaction (with NUDT18, Q6ZVK8) captured in a
      proteome-scale high-throughput yeast two-hybrid interactome map. "protein binding"
      is uninformative and does not describe a characterized molecular function of DUT.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Per curation guidelines, bare "protein binding" is uninformative. This is a
      high-throughput Y2H interactome hit (with NUDT18) with no established functional
      consequence for dUTPase. Retained (not removed) as an experimental IPI but marked
      as over-annotated because it does not capture a core molecular function.
    supported_by:
    - reference_id: PMID:16189514
      supporting_text: >-
        we tested pairwise interactions among the products of approximately 8,100
        currently available Gateway-cloned open reading frames and detected
        approximately 2,800 interactions
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19060904
  qualifier: enables
  review:
    summary: >-
      Second IntAct-curated binary interaction record (with NUDT18, Q6ZVK8) from the
      empirical binary interactome-mapping framework study. Again bare "protein binding".
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Bare "protein binding" from a high-throughput binary interactome study; not an
      informative molecular function for DUT. Retained as experimental IPI but marked
      over-annotated.
    supported_by:
    - reference_id: PMID:19060904
      supporting_text: >-
        we predict that the entire human interactome, excluding splice variant
        complexity, contains approximately 74,000-200,000 binary biophysical interactions
- term:
    id: GO:0001889
    label: liver development
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: involved_in
  review:
    summary: >-
      Ensembl-Compara electronic transfer from the rat ortholog (P70583). There is no
      human evidence that dUTPase has a specific role in liver development; this is a
      peripheral, ortholog-projected annotation.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      dUTPase is a ubiquitous housekeeping nucleotide-metabolism enzyme; "liver
      development" is an over-specific ortholog-projected process not reflecting DUT's
      core function. Marked over-annotated rather than removed, since it derives from
      an experimentally verified rat annotation transferred by Ensembl.
    supported_by:
    - reference_id: PMID:1325640
      supporting_text: >-
        All of these mRNAs are present in a variety of human tissues
- term:
    id: GO:0030547
    label: signaling receptor inhibitor activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: enables
  review:
    summary: >-
      Ensembl-Compara electronic transfer from the rat ortholog. Reflects the reported
      "By similarity" moonlighting property in which dUTPase can inhibit PPAR by binding
      its N-terminus and blocking PPAR:RXR dimerization.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      This corresponds to a peripheral moonlighting activity (PPAR inhibition) inferred
      by similarity, not the core enzymatic function of DUT, and it derives from an
      ortholog projection rather than direct human evidence. Not core; marked
      over-annotated rather than removed.
    supported_by:
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: >-
        preventing the latter's dimerization with retinoid X receptor
- term:
    id: GO:0032556
    label: pyrimidine deoxyribonucleotide binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: enables
  review:
    summary: >-
      Ensembl-Compara electronic transfer of pyrimidine deoxyribonucleotide binding.
      dUTPase does bind the pyrimidine deoxyribonucleotide dUTP as its substrate.
    action: ACCEPT
    reason: >-
      Correct: dUTPase specifically binds the pyrimidine deoxyribonucleotide dUTP
      (structurally characterized substrate binding), consistent with this term.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        complexes with the uracil-containing deoxyribonucleotides, dUMP, dUDP and dUTP
- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: enables
  review:
    summary: >-
      Ensembl-Compara electronic transfer of identical protein binding, consistent with
      dUTPase functioning as a homotrimer (self-association).
    action: KEEP_AS_NON_CORE
    reason: >-
      dUTPase is an obligate homotrimer, so "identical protein binding" is factually
      correct as a structural correlate of oligomerization; however it is not the
      informative core molecular function (which is the catalytic dUTP diphosphatase
      activity). Kept as non-core.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        each subunit of the dUTPase trimer folds into an eight-stranded jelly-roll
        beta barrel
- term:
    id: GO:0042975
    label: peroxisome proliferator activated receptor binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: enables
  review:
    summary: >-
      Ensembl-Compara electronic transfer of PPAR binding, corresponding to the reported
      "By similarity" moonlighting interaction in which dUTPase binds PPAR via its
      N-terminus.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Peripheral moonlighting interaction inferred by similarity/ortholog projection,
      not the core function of DUT and without direct human experimental support in the
      cited evidence. Marked over-annotated rather than removed.
    supported_by:
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: >-
        Inhibits peroxisome proliferator-
- term:
    id: GO:0043254
    label: regulation of protein-containing complex assembly
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: involved_in
  review:
    summary: >-
      Ensembl-Compara electronic transfer of "regulation of protein-containing complex
      assembly", plausibly relating to the reported inhibition of PPAR:RXR dimerization
      by dUTPase.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      This is an indirect, ortholog-projected process linked to the peripheral PPAR
      moonlighting activity, not the core nucleotide-metabolism function of DUT. Marked
      over-annotated rather than removed.
    supported_by:
    - reference_id: file:human/DUT/DUT-uniprot.txt
      supporting_text: >-
        preventing the latter's dimerization with retinoid X receptor
- term:
    id: GO:0005654
    label: nucleoplasm
  evidence_type: IDA
  original_reference_id: GO_REF:0000052
  qualifier: located_in
  review:
    summary: >-
      Human Protein Atlas immunofluorescence localization to the nucleoplasm,
      consistent with the nuclear isoform (DUT-N).
    action: ACCEPT
    reason: >-
      Nucleoplasmic localization is consistent with the experimentally established
      nuclear localization of the DUT-N isoform and with Reactome's placement of the
      major isoform in the nucleoplasm.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the lower molecular weight form of dUTPase (DUT-N) is associated with the
        nucleus
- term:
    id: GO:0004170
    label: dUTP diphosphatase activity
  evidence_type: EXP
  original_reference_id: PMID:17880943
  qualifier: enables
  review:
    summary: >-
      Direct experimental (structural/mechanistic) demonstration of dUTPase catalysis
      via the crystal structure of the enzyme:substrate-analog:Mg complex.
    action: ACCEPT
    reason: >-
      Core, directly demonstrated molecular function of DUT, established by structural
      and equilibrium-binding characterization of the catalytic mechanism.
    supported_by:
    - reference_id: PMID:17880943
      supporting_text: >-
        We determined the crystal structure of the
        enzyme:alpha,beta-imino-dUTP:Mg complex and performed equilibrium binding
        experiments in solution.
- term:
    id: GO:0004170
    label: dUTP diphosphatase activity
  evidence_type: EXP
  original_reference_id: PMID:8805593
  qualifier: enables
  review:
    summary: >-
      Direct experimental demonstration of dUTP diphosphatase activity via the first
      atomic-resolution structure of human dUTPase in complex with uracil-containing
      deoxyribonucleotides.
    action: ACCEPT
    reason: >-
      Core, directly demonstrated molecular function; the structure captures the
      enzyme bound to dUMP/dUDP/dUTP and defines the catalytic active site.
    supported_by:
    - reference_id: PMID:8805593
      supporting_text: >-
        dUTPase hydrolyzes dUTP to dUMP and pyrophosphate
- term:
    id: GO:0005634
    label: nucleus
  evidence_type: EXP
  original_reference_id: PMID:9070952
  qualifier: located_in
  review:
    summary: >-
      Experimental subcellular localization to the nucleus (nuclear isoform), cited by
      UniProt for the DUT-N isoform.
    action: ACCEPT
    reason: >-
      Nuclear localization of the nuclear dUTPase isoform is experimentally established;
      accept and defer to the curator who assessed the full text. The cached record is
      abstract-only, but UniProt cites this reference for the isoform-2 nuclear location.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the lower molecular weight form of dUTPase (DUT-N) is associated with the
        nucleus
- term:
    id: GO:0005739
    label: mitochondrion
  evidence_type: EXP
  original_reference_id: PMID:9070952
  qualifier: located_in
  review:
    summary: >-
      Experimental subcellular localization to the mitochondrion (mitochondrial isoform
      DUT-M), cited by UniProt for isoform 3.
    action: ACCEPT
    reason: >-
      Mitochondrial localization of the DUT-M isoform is experimentally established;
      accept and defer to the curator. Independently supported by high-confidence
      mitochondrial proteomics.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the higher molecular weight species (DUT-M) fractionates with the mitochondria
- term:
    id: GO:0005739
    label: mitochondrion
  evidence_type: HTP
  original_reference_id: PMID:34800366
  qualifier: located_in
  review:
    summary: >-
      High-throughput identification in a quantitative high-confidence human
      mitochondrial proteome (MitoCoP), consistent with the mitochondrial DUT-M isoform.
    action: ACCEPT
    reason: >-
      Corroborates the experimentally established mitochondrial localization of the
      DUT-M isoform via a high-confidence mitochondrial proteomics resource.
    supported_by:
    - reference_id: PMID:34800366
      supporting_text: >-
        defined a mitochondrial high-confidence proteome of >1,100 proteins (MitoCoP)
- term:
    id: GO:0004170
    label: dUTP diphosphatase activity
  evidence_type: IDA
  original_reference_id: PMID:8631816
  qualifier: enables
  review:
    summary: >-
      Direct assay of dUTP diphosphatase activity on purified human dUTPase (both
      nuclear and mitochondrial forms show identical dUTP binding).
    action: ACCEPT
    reason: >-
      Core, directly demonstrated molecular function. dUTPase was purified from HeLa
      cells and both isoforms exhibited identical dUTP binding (KM ~2.5 uM).
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        both forms of dUTPase exhibited identical binding characteristics for dUTP
- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IDA
  original_reference_id: PMID:8631816
  qualifier: located_in
  review:
    summary: >-
      Direct demonstration (cellular fractionation / isopycnic density centrifugation)
      that the nuclear dUTPase isoform (DUT-N) localizes to the nucleus.
    action: ACCEPT
    reason: >-
      Directly demonstrated nuclear localization of the DUT-N isoform.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the lower molecular weight form of dUTPase (DUT-N) is associated with the
        nucleus
- term:
    id: GO:0006231
    label: dTMP biosynthetic process
  evidence_type: IDA
  original_reference_id: PMID:10952785
  qualifier: acts_upstream_of_or_within
  review:
    summary: >-
      dUTPase supplies dUMP and maintains low dUTP, acting upstream of thymidylate
      synthase in de novo dTMP biosynthesis; dUTPase expression/activity modulates the
      dUTP pool and sensitivity to thymidylate synthase inhibition.
    action: ACCEPT
    reason: >-
      dUTPase is functionally upstream of dTMP biosynthesis: it produces the TYMS
      substrate dUMP and limits uracil misincorporation when dTTP is depleted. The
      acts_upstream_of_or_within qualifier is appropriate.
    supported_by:
    - reference_id: PMID:10952785
      supporting_text: >-
        dUTPase catalyses the hydrolysis of dUTP to dUMP, thereby maintaining low
        intracellular dUTP.
- term:
    id: GO:0006231
    label: dTMP biosynthetic process
  evidence_type: IMP
  original_reference_id: PMID:15322254
  qualifier: acts_upstream_of_or_within
  review:
    summary: >-
      siRNA knockdown of dUTPase perturbs the dUTP pool and sensitizes cancer cells to
      the thymidylate synthase inhibitor FUdR, functionally linking dUTPase to dTMP
      biosynthesis / thymineless pathways.
    action: ACCEPT
    reason: >-
      Loss-of-function (siRNA) evidence places dUTPase upstream of dTMP biosynthesis and
      the thymidylate-synthase pathway; acts_upstream_of_or_within is appropriate.
    supported_by:
    - reference_id: PMID:15322254
      supporting_text: >-
        Suppression of dUTPase in SW620 and MCF-7 cells resulted in a significant
        enhancement in dUTP pool expansion after TS inhibition.
- term:
    id: GO:0005634
    label: nucleus
  evidence_type: IDA
  original_reference_id: PMID:8631816
  qualifier: is_active_in
  review:
    summary: >-
      The nuclear dUTPase isoform (DUT-N) is active in the nucleus, where it depletes
      dUTP to protect replicating DNA from uracil misincorporation.
    action: ACCEPT
    reason: >-
      is_active_in nucleus is appropriate: the catalytically active DUT-N isoform is
      localized to the nucleus, the compartment where prevention of uracil
      misincorporation into replicating DNA is most relevant.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the lower molecular weight form of dUTPase (DUT-N) is associated with the
        nucleus
- term:
    id: GO:0006231
    label: dTMP biosynthetic process
  evidence_type: IDA
  original_reference_id: PMID:10952785
  qualifier: involved_in
  review:
    summary: >-
      dUTPase is involved in de novo dTMP biosynthesis by producing dUMP and keeping
      dUTP low; dUTPase activity modulates sensitivity to thymidylate synthase inhibition.
    action: ACCEPT
    reason: >-
      Involvement in dTMP biosynthesis is well supported: dUTPase furnishes the dUMP
      substrate for thymidylate synthase and its activity governs the dUTP:dTTP balance.
    supported_by:
    - reference_id: PMID:10952785
      supporting_text: >-
        dUTPase catalyses the hydrolysis of dUTP to dUMP, thereby maintaining low
        intracellular dUTP.
- term:
    id: GO:0006231
    label: dTMP biosynthetic process
  evidence_type: IDA
  original_reference_id: PMID:15322254
  qualifier: involved_in
  review:
    summary: >-
      dUTPase involvement in dTMP biosynthesis / thymidylate metabolism, supported by
      the effect of dUTPase suppression on dUTP pools and thymidylate-synthase-inhibitor
      sensitivity.
    action: ACCEPT
    reason: >-
      Involved_in dTMP biosynthetic process is supported by loss-of-function evidence
      linking dUTPase to the dUTP/dTTP balance and thymidylate synthesis pathway.
    supported_by:
    - reference_id: PMID:15322254
      supporting_text: >-
        dUTPase, which eliminates dUTP from the DNA biosynthetic pathway, opposes
        uracil misincorporation
- term:
    id: GO:0003723
    label: RNA binding
  evidence_type: HDA
  original_reference_id: PMID:22658674
  qualifier: enables
  review:
    summary: >-
      Detected as an mRNA-binding protein in a HeLa "interactome capture" (UV
      crosslinking) proteomic screen. This is a high-throughput moonlighting/incidental
      RNA-association finding, not a characterized function of dUTPase.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      RNA binding was reported only from a global mRNA-interactome capture screen; there
      is no evidence that RNA binding is a core or physiologically important function of
      dUTPase (a nucleotide-hydrolase). Treated as non-core moonlighting and marked
      over-annotated. Retained (not removed) as a high-throughput experimental result.
    supported_by:
    - reference_id: PMID:22658674
      supporting_text: >-
        We identify 860 proteins that qualify as RBPs by biochemical and statistical
        criteria
- term:
    id: GO:0070062
    label: extracellular exosome
  evidence_type: HDA
  original_reference_id: PMID:20458337
  qualifier: located_in
  review:
    summary: >-
      Detected by mass spectrometry in the proteome of B-cell-derived exosomes. This is
      a high-throughput proteomic survey; dUTPase is an intracellular nucleotide-
      metabolism enzyme and is not a functional exosomal component.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Extracellular exosome detection reflects incidental presence in a bulk exosome
      proteome, not a functionally meaningful localization for a nuclear/mitochondrial
      housekeeping enzyme. Marked over-annotated rather than removed.
    supported_by:
    - reference_id: PMID:20458337
      supporting_text: >-
        we first analyzed the total proteome of highly purified B cell-derived exosomes
        using sensitive and accurate mass spectrometry (MS), and identified 539 proteins
- term:
    id: GO:0005654
    label: nucleoplasm
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-73666
  qualifier: located_in
  review:
    summary: >-
      Reactome places the major (nuclear) dUTPase isoform in the nucleoplasm, where it
      catalyzes hydrolysis of dUTP to dUMP and pyrophosphate.
    action: ACCEPT
    reason: >-
      Consistent with the experimentally established nuclear localization of the DUT-N
      isoform and with the HPA nucleoplasm IDA annotation.
    supported_by:
    - reference_id: Reactome:R-HSA-73666
      supporting_text: >-
        The major one, annotated here, is localized to the nucleoplasm
- term:
    id: GO:0004170
    label: dUTP diphosphatase activity
  evidence_type: TAS
  original_reference_id: PMID:8631816
  qualifier: enables
  review:
    summary: >-
      Traceable author statement of dUTP diphosphatase activity for human dUTPase.
    action: ACCEPT
    reason: >-
      Redundant with the direct experimental annotations for the same term; correct core
      molecular function.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        both forms of dUTPase exhibited identical binding characteristics for dUTP
- term:
    id: GO:0005634
    label: nucleus
  evidence_type: TAS
  original_reference_id: PMID:8631816
  qualifier: located_in
  review:
    summary: >-
      Traceable author statement of nuclear localization for the DUT-N isoform.
    action: ACCEPT
    reason: >-
      Redundant with the IDA/EXP nucleus annotations; correct localization of the
      nuclear isoform.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the lower molecular weight form of dUTPase (DUT-N) is associated with the
        nucleus
- term:
    id: GO:0005739
    label: mitochondrion
  evidence_type: TAS
  original_reference_id: PMID:8631816
  qualifier: located_in
  review:
    summary: >-
      Traceable author statement of mitochondrial localization for the DUT-M isoform.
    action: ACCEPT
    reason: >-
      Redundant with the EXP/HTP mitochondrion annotations; correct localization of the
      mitochondrial isoform.
    supported_by:
    - reference_id: PMID:8631816
      supporting_text: >-
        the higher molecular weight species (DUT-M) fractionates with the mitochondria
- term:
    id: GO:0006139
    label: nucleobase-containing compound metabolic process
  evidence_type: TAS
  original_reference_id: PMID:1325640
  qualifier: involved_in
  review:
    summary: >-
      Traceable author statement placing dUTPase in nucleotide/nucleobase-containing
      compound metabolism. This is a very broad parent process.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Correct but extremely general; the specific dUTP catabolic / dUMP biosynthetic /
      dTMP biosynthetic process annotations already capture DUT's role at the
      appropriate level of granularity. Marked over-annotated as a too-broad term.
    supported_by:
    - reference_id: PMID:1325640
      supporting_text: >-
        dUTPase may generally perform an essential role in DNA replication
- term:
    id: GO:0006260
    label: DNA replication
  evidence_type: TAS
  original_reference_id: PMID:1325640
  qualifier: involved_in
  review:
    summary: >-
      Traceable author statement associating dUTPase with DNA replication. dUTPase
      supports faithful replication by keeping dUTP low (preventing uracil
      misincorporation) and supplying dUMP for dTTP, but it is not part of the
      replication machinery itself.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      dUTPase influences the fidelity of DNA replication indirectly, by controlling the
      deoxyribonucleotide pool, rather than acting in the DNA replication process per se.
      "DNA replication" (involved_in) over-states its direct role; the causal
      contribution is better captured by its nucleotide-metabolism annotations. Marked
      over-annotated rather than removed (traceable author statement).
    supported_by:
    - reference_id: PMID:1325640
      supporting_text: >-
        dUTPase may generally perform an essential role in DNA replication
core_functions:
- description: >-
    Mg2+-dependent dUTP diphosphatase (dUTPase) that hydrolyses dUTP to dUMP and
    inorganic diphosphate, catabolising dUTP and supplying dUMP for de novo
    thymidylate (dTMP) biosynthesis while preventing uracil misincorporation into DNA.
  molecular_function:
    id: GO:0004170
    label: dUTP diphosphatase activity
  directly_involved_in:
  - id: GO:0046081
    label: dUTP catabolic process
  supported_by:
  - reference_id: PMID:8805593
    supporting_text: >-
      dUTPase hydrolyzes dUTP to dUMP and pyrophosphate, simultaneously reducing dUTP
      levels and providing the dUMP for dTTP biosynthesis.
  - reference_id: PMID:17880943
    supporting_text: >-
      We determined the crystal structure of the
      enzyme:alpha,beta-imino-dUTP:Mg complex
- description: >-
    Production of dUMP (the immediate thymidylate-synthase substrate) as the direct
    product of dUTP hydrolysis, feeding de novo dTMP biosynthesis in the nucleus and
    cytosol.
  molecular_function:
    id: GO:0004170
    label: dUTP diphosphatase activity
  directly_involved_in:
  - id: GO:0006226
    label: dUMP biosynthetic process
  locations:
  - id: GO:0005634
    label: nucleus
  - id: GO:0005739
    label: mitochondrion
  supported_by:
  - reference_id: Reactome:R-HSA-73666
    supporting_text: >-
      this reaction depletes the supply of dUTP, preventing its incorporation into DNA,
      while generating dUMP, the immediate precursor of thymidine nucleotides.
  - reference_id: PMID:10952785
    supporting_text: >-
      dUTPase catalyses the hydrolysis of dUTP to dUMP, thereby maintaining low
      intracellular dUTP.
references:
- id: GO_REF:0000002
  title: Gene Ontology annotation through association of InterPro records with GO
    terms
  findings: []
- 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: GO_REF:0000052
  title: Gene Ontology annotation based on curation of immunofluorescence data
  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:10952785
  title: Deoxyuridine triphosphatase (dUTPase) expression and sensitivity to the thymidylate
    synthase (TS) inhibitor ZD9331.
  findings: []
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: >-
      Abstract/full-text verified; supports dUTPase = hydrolysis of dUTP to dUMP and
      its upstream role relative to thymidylate synthase.
- id: PMID:1325640
  title: 'Human dUTP pyrophosphatase: cDNA sequence and potential biological importance
    of the enzyme.'
  findings: []
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: >-
      Original human dUTPase cDNA cloning; supports family assignment and essential role
      in DNA replication; the DNA-replication/broad-metabolism GO annotations from this
      TAS are over-broad.
- id: PMID:15322254
  title: Small interfering RNA-mediated suppression of dUTPase sensitizes cancer cell
    lines to thymidylate synthase inhibition.
  findings: []
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Loss-of-function evidence placing dUTPase upstream of dTMP biosynthesis / TS
      inhibition via dUTP-pool control.
- id: PMID:16189514
  title: Towards a proteome-scale map of the human protein-protein interaction network.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: >-
      Proteome-scale HT-Y2H interactome; source of a bare "protein binding" IPI (with
      NUDT18). No functional consequence for dUTPase.
- id: PMID:17880943
  title: Active site closure facilitates juxtaposition of reactant atoms for initiation
    of catalysis by human dUTPase.
  findings: []
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Structure of the human dUTPase:imino-dUTP:Mg complex; supports dUTP diphosphatase
      activity and Mg2+ binding.
- id: PMID:19060904
  title: An empirical framework for binary interactome mapping.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: >-
      Binary interactome-mapping framework; source of a second bare "protein binding"
      IPI. Not informative for dUTPase function.
- id: PMID:20458337
  title: MHC class II-associated proteins in B-cell exosomes and potential functional
    implications for exosome biogenesis.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: >-
      B-cell exosome proteome MS survey; source of the extracellular-exosome HDA. Not a
      functional localization for dUTPase.
- id: PMID:22658674
  title: Insights into RNA biology from an atlas of mammalian mRNA-binding proteins.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: >-
      mRNA interactome-capture atlas; source of the RNA-binding HDA. Treated as non-core
      moonlighting.
- id: PMID:34800366
  title: Quantitative high-confidence human mitochondrial proteome and its dynamics
    in cellular context.
  findings: []
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: >-
      High-confidence mitochondrial proteome (MitoCoP); corroborates mitochondrial
      localization of the DUT-M isoform.
- id: PMID:8631816
  title: Characterization of distinct nuclear and mitochondrial forms of human deoxyuridine
    triphosphate nucleotidohydrolase.
  findings: []
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Defines the nuclear (DUT-N) and mitochondrial (DUT-M) isoforms, their identical
      dUTP binding, and their distinct localizations. Key reference for activity and
      localization.
- id: PMID:8805593
  title: 'Human dUTP pyrophosphatase: uracil recognition by a beta hairpin and active
    sites formed by three separate subunits.'
  findings: []
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      First atomic-resolution structure of human dUTPase; defines catalysis, homotrimer,
      Mg2+ cofactor, dUTP->dUMP+PPi, and dUTP:dTTP-ratio rationale.
- id: PMID:9070952
  title: Assignment of the human dUTPase gene (DUT) to chromosome 15q15-q21. 1 by
    fluorescence in situ hybridization.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: >-
      Chromosomal assignment (15q15-q21.1). Cited by UniProt for isoform localization;
      cached record is abstract-only.
- id: Reactome:R-HSA-73666
  title: hydrolysis of  2'-deoxyuridine 5'-triphosphate to form  2'-deoxyuridine 5'-phosphate
  findings: []
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Reactome reaction for dUTP -> dUMP + PPi; nucleoplasmic major isoform; homotrimer.
- id: file:human/DUT/DUT-uniprot.txt
  title: UniProtKB entry P33316 (DUT_HUMAN)
  findings: []
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      UniProt curated record used for pathway (dUMP biosynthesis; dUMP from dCTP),
      dUTPase family assignment, and the By-similarity PPAR moonlighting function.
proposed_new_terms: []
suggested_questions:
- question: >-
    Do the nuclear (DUT-N) and mitochondrial (DUT-M) isoforms have non-redundant
    physiological roles, or does one compartment's activity suffice for genome
    integrity under normal conditions?
- question: >-
    Is the reported PPAR-inhibition / signaling-receptor-inhibitor moonlighting
    function present and physiologically relevant in humans, or is it specific to the
    rodent ortholog from which it was projected?
suggested_experiments:
- description: >-
    Isoform-specific rescue of DUT-null cells with nuclear-only vs mitochondrial-only
    dUTPase to test compartmental requirements for preventing uracil misincorporation.
- description: >-
    Quantify dUTP:dTTP pools and uracil-DNA content in cells carrying the BMFDMS
    variants (Y142C, R173W, Y227C) to link catalytic impairment to the disease
    phenotype.