Gene Ontology annotation through association of InterPro records with GO terms
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The route behind the GO:0000287 and GO:0051725 rows, and behind GO:0003875 in the non-mammalian ARH2 orthologues. Its input is IPR012108, whose interpro2go entry maps to exactly those three terms and whose reviewed membership is 4 ARH1 proteins and 7 Swiss-Prot-declared inactive ARH2 proteins.
Manual transfer of experimentally-verified manual GO annotation data to orthologs by curator judgment of sequence similarity
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The route behind the three ISS rows, all citing UniProtKB:Q6AZR2 (ARHL1_XENLA), a reviewed 1:1 ortholog at 74.6% aligned identity. Correct WITH/FROM entity type for ISS. The limitation is not in the transfer but in the donor set: the same Xenopus gene also has Xenbase-curated annotations on TrEMBL accessions that this route never sees.
Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping, accompanied by conservative changes to GO terms applied by UniProt
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The route behind the second GO:0030017 row. Its input is SubCell SL-0313, which comes from this entry's SUBCELLULAR LOCATION line, itself ECO:0000250|UniProtKB:Q6AZR2 - so this row and the ISS GO:0030017 row are one observation reaching GOA twice.
Combined Automated Annotation using Multiple IEA Methods
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The route behind the GO:0003875 row in the five mammalian ARH2 orthologues. Its WITH/FROM names ARBA:ARBA00088955, whose full condition set - fetched from the UniProt ARBA endpoint - is FunFam id 1.10.4080.10:FF:000002 AND taxon Mammalia. The Mammalia clause is directly visible in GOA as the GO_REF:0000120 versus GO_REF:0000002 split between the mammalian and non-mammalian orthologues.
The cardiac-restricted protein ADP-ribosylhydrolase-like 1 is essential for heart chamber outgrowth and acts on muscle actin filament assembly.
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The experimental root of all three ISS rows. Morpholino knockdown of all Adprhl1 variants in Xenopus laevis blocks striated myofibril assembly and prevents ventricle outgrowth, without affecting cardiomyocyte number, calcium signal propagation or myofibrillar gene expression.
"Morpholino oligonucleotide-mediated knockdown of all Adprhl1 variants inhibits striated myofibril assembly and prevents outgrowth of the ventricle."
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Structural modelling on the ADPRH crystal structure 3HFW identifies the specific losses in the ADPRHL1 active site, listing them in ADPRH numbering.
"Three of the four critical aspartates of ADPRH have been lost in mammalian ADPRHL1 (hADPRH D56, D302, D304 lost, only D55 is conserved), suggesting that ADPRHL1 cation binding may be compromised."
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The same modelling records a hedged possibility that a different aspartate could coordinate metal, which is why the GO:0000287 verdict is argued from the annotation's derivation rather than from a flat claim that ADPRHL1 binds no metal.
"Of particular note is the aspartate, D100, that could provide an alternative coordination site for metals but whose presence interferes with the phosphate groups of forcibly docked ADP."
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The source of the sarcomere localisation, obtained with over-expressed N-terminally tagged protein.
"Recombinant Adprhl1 can localize to stripes adjacent to the Z-disc"
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The caveat on that localisation, from the same study - the endogenous protein could not be detected in situ, so no endogenous localisation exists in any species.
"immunocytochemistry of hearts using the same antibody was not sufficiently sensitive to detect endogenous Adprhl1 protein in situ"
Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site.
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The genetic demonstration that ADPRHL1's function is cleft-dependent and catalysis-independent: CRISPR-induced in-frame deletions of one to four residues from the di-arginine loop at the centre of the vestigial ADP-ribosylhydrolase site abolish ventricular myofibril assembly.
"Mutant alleles encode discrete loss of 1, 3 or 4 amino acids from a di-arginine (Arg271-Arg272) containing peptide loop at the centre of the ancestral ADP-ribosylhydrolase site."
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The authors' own summary of what that means for a pseudoenzyme.
"Thus despite lacking catalytic activity, it is the modified (adenosine-ribose) substrate binding cleft of Adprhl1 that fulfils an essential role during heart formation."
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The structural claim that this review's alignment independently reproduces from UniProt features alone - ADPRH's substrate-binding Ser269 and Ser270 map onto ADPRHL1's Arg271 and Arg272.
"The critical Adprhl1 deletion covers the exact structural position where in the active enzyme Adprh, two adjacent serines that support adenosine-ribose substrate binding are located."
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The secondary attribution for the biochemical negative, which this review anchors independently to PMID:17075046 and PMID:36497109.
"The sequence changes suggest binding of ADP-ribosylated protein substrates and cation-mediated catalysis are both abolished in Adprhl1 and biochemical assays have confirmed the lack of any comparable enzymatic activity"
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The only published mammalian allele, and the reason it does not test the requirement - it is not a null.
"Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected"
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The published identity figures used as external checks that this review's alignment method is sane; measured 46.6% and 74.6% against 46% and 75%.
"the 357 amino acid ADPRH and 354 aa ADPRHL1 share 46% sequence identity"
The pseudoenzyme ADPRHL1 affects cardiac function by regulating the ROCK pathway.
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The only functional experiment ever performed on the human protein. CRISPR knockout in H9 embryonic stem cell-derived cardiomyocytes; the cells still differentiate and still express cardiac genes, but adhere abnormally and show perturbed calcium handling and electrophysiology.
"Our results demonstrated that ADPRHL1-deficient cardiomyocytes exhibited abnormal adhesion, calcium transients and electrophysiological activity."
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The mechanism and its direction - knockout reduces focal adhesion formation via excessive ROCK-myosin II activity, so wild-type ADPRHL1 promotes focal adhesion assembly.
"ADPRHL1 deficiency disrupted the formation of focal adhesions in cardiomyocytes by excessively upregulating the ROCK"
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The focal adhesion component measurements underlying that conclusion.
"suggesting that ADPRHL1 deficiency disrupts the FAs in these cells"
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A second, independent focal adhesion marker measured in the same cells.
"the expression of Paxillin, a structural protein in FAs, in KO-CM was markedly reduced"
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Independent human support for the cardiac myofibril assembly annotation that GOA currently holds only by ISS from frog.
"the arrangement of sarcomeres was less regular"
The 39-kDa poly(ADP-ribose) glycohydrolase ARH3 hydrolyzes O-acetyl-ADP-ribose, a product of the Sir2 family of acetyl-histone deacetylases.
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The direct in vitro measurement that recombinant ARH2 has no activity, made alongside ARH1 and ARH3 in the same assay. This is the single most decision-relevant fact for this gene's molecular function rows, and it is in a paper titled for a paralogue.
"The rate of O-acetyl-ADP-ribose hydrolysis by recombinant ARH3 was 250-fold that observed with ARH1; ARH2 and poly(ADP-ribose) glycohydrolase were inactive."
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The mutagenesis that anchors this review's claim that aspartate-to-asparagine at a vicinal catalytic aspartate is inactivating rather than conservative - the same substitution ADPRHL1 carries at the position corresponding to ADPRH Asp56.
"hydrolysis of O-acetyl-ADP-ribose was abolished by replacement of the vicinal aspartates at positions 77 and 78 of ARH3 with asparagine"
ARH Family of ADP-Ribose-Acceptor Hydrolases.
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The family review's statement of ARH2's enzymatic status, with the specific sequence motifs whose loss explains it. The ARH1 motifs quoted, 54-SDDT-57 and 302-DSDS-305, correspond exactly to the UniProt-annotated magnesium ligands used as the reference set in this review's residue census.
"ARH2 (so far) does not exhibit any enzymatic activities [70], consistent with differences in primary sequences from critical residues found in ARH1 (e.g., 54-SDDT-57, 302-DSDS-305)"
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The statement bearing specifically on the reaction that GO:0003875 and GO:0051725 name.
"ARH2 does not appear to be responsible for hydrolysis of ADP-ribose-acceptor linkage."
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The authors' own caveat, recorded because it is the reason no replacement hydrolase term is proposed and the reason this review does not claim ADPRHL1 is biochemically inert.
"However, the lack of ARH2 catalytic activity may be the result of the use of model substrates."
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The expression pattern and the biological role attributed to ARH2 by the family's own reviewers.
"ARH2 is a cytoplasmic protein expressed primarily in heart and skeletal muscle and may be involved in cardiac myofibril assembly and cardiac chamber outgrowth"
A Recurrent ADPRHL1 Germline Mutation Activates PARP1 and Confers Prostate Cancer Risk in African American Families.
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A recurrent germline p.Asp78Val allele segregating in African-American prostate cancer families; the mutant activates PARP1 and increases the DNA damage response, an effect reversed by olaparib.
"the ADPRHL1 mutant activates PARP1"
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Ectopic wild-type ADPRHL1 restrains proliferation in prostate cancer cells. Suggestive of a link to cellular ADP-ribosylation but not evidence that ADPRHL1 itself acts on ADP-ribose, so no annotation is proposed from it.
"expression of the wild-type ADPRHL1 in prostate cancer cells suppressed cell proliferation and oncogenesis"
Inhibition of ARH2 by pH/ROS-responsive nanosystem for improved lung adenocarcinoma immunochemotherapy.
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siRNA knockdown of ARH2 delivered by nanoparticle in lung adenocarcinoma models; ARH2 is reported to promote M2 macrophage polarisation via FPR2, PI3K and AKT. A different tissue and the opposite direction from the prostate work, and rated low confidence by the provider that surfaced it.
"ARH2 promotes M2 macrophage polarization and suppresses immune responses by regulating the FPR2/PI3K/AKT signaling pathway"
Histone deacetylase 4 deletion broadly affects cardiac epigenetic repression and regulates transcriptional susceptibility via H3K9 methylation.
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Identifies the Adprhl1 promoter as a target of HDAC4, MEF2 and SUV39H1 repression in cardiomyocytes. This concerns regulation of the gene, not a function of the protein, and yields no annotation.
Identification and characterization of a mammalian 39-kDa poly(ADP-ribose) glycohydrolase.
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The reference both Xenopus papers cite for ARH2's lack of comparable enzymatic activity. Cached abstract-only, and the abstract does not itself mention ARH2, so no claim in this review rests on it - the biochemical negative is anchored to PMID:17075046 and PMID:36497109 instead.
The family of toxin-related ecto-ADP-ribosyltransferases in humans and the mouse.
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UniProt's reference 1 for the ADPRHL1 isoform-1 mRNA. It surveys the ecto-ADP-ribosyltransferase (ARTC) family, which is a different family from the ARH hydrolases, and is the most likely origin of the word transferase in Q8NDY3's recommended name.
Affinage mechanistic annotation for ADPRHL1 (human)
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The provider record used as a starting lead, recorded for provider evaluation rather than cited as evidence. Its narrative is broadly accurate and all six of its citations were individually verified to concern ADPRHL1 or ARH2 - which is not a given, since on the sibling gene ADPRH the same provider returned citations resolving to papers about entirely different proteins.
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Its recall, however, missed every source that decides this review's molecular function verdicts: PMID:17075046 (the only direct measurement that ARH2 is inactive, titled for ARH3), PMID:36497109 (the family review that states the negative across the assay panel and supplies the model-substrate caveat) and PMID:16278211 (the reference the primary literature cites for ARH2's inactivity). All three were found by searching the family rather than the gene symbol.
UniProtKB entry Q8NDY3 (ARHL1_HUMAN), Inactive ADP-ribosyltransferase ARH2
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The curated record states the catalytic loss twice - once in FUNCTION and once in CAUTION - and its feature table contains zero BINDING and zero ACT_SITE features. ADPRH (P54922) has 14 such features spanning 20 distinct residue positions and ADPRS (Q9NX46) 14 spanning 17; ADPRHL1 has none at all.
"lacks the metal-binding and substrate-binding residues"
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The FUNCTION line's statement of the measured negative, attributed by similarity to the rat and mouse entries.
"showing no activity against O-acetyl-ADP-ribose"
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The input to the SubCell IEA row, itself ECO:0000250|UniProtKB:Q6AZR2 and therefore not independent of the ISS row.
"SUBCELLULAR LOCATION: Cytoplasm, myofibril, sarcomere"
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Corroborates the QuickGO finding that this gene has no IBA annotation, which is a deliberate PAINT judgement rather than a coverage gap.
"0 GO annotations based on evolutionary models."
ADPRHL1 (ARH2, Q8NDY3): catalytic-site census against its active paralogues
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The identity-matched control that turns the residue observation into an argument. Dictyostelium ADPRH, a genuine ARH1 at 48.4% identity - the same distance as ADPRHL1's 42.6-47.7% - keeps 13 of 20 ligand sites and all six magnesium donors, while every ADPRHL1 orthologue keeps 6-7 sites and 2-3 donors.
"Retention therefore is not tracking sequence distance here."
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The low-identity active controls pointing the same way from the other side, and the reason the result cannot be explained by sequence divergence alone.
"Two low-identity active enzymes point the same way: DraG at 27.5% keeps 9, and the"
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The independent reproduction of the sibling ADPRH review's panel, split into the objective channel (residue and mapped position) and the metric channel (class label).
"identical on all 25 calls"