Naked-mole-rat cGAS is a DNA-binding nucleotidyltransferase with conserved machinery for synthesizing the second messenger 2β²3β²-cGAMP from ATP and GTP. The mammalian cGAMP-STING innate immune pathway is inferred to be retained. In the nucleus, naked-mole-rat cGAS promotes homologous recombination repair: species-specific C-terminal changes weaken TRIM41-mediated ubiquitination and p97 engagement, prolong chromatin retention after DNA damage, and enhance FANCI-RAD50 association and RAD50 recruitment. This repair-promoting behavior differs from the suppressive effect of human and mouse cGAS. The magnitude and regulation of cGAMP and interferon output have not been measured for this species in the inspected evidence.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0002218 activation of innate immune response | IEA GO_REF:0000118 | ACCEPT | Summary: Retain the conserved cGAS contribution to innate immune activation. Reason: Innate immune activation is the inherited signaling role of cGAMP production upstream of STING; target repair-specialization evidence does not negate that separate core signaling branch. The curated ancestral cGAS inference is supported by the intact catalytic triad, zinc thumb and nucleotide-binding machinery in A0AAX6RS70 and mammalian cGAMP-STING experiments. PMID:41066557 establishes reversal of the nuclear homologous-recombination phenotype, not loss of innate immune signaling. Prolonged chromatin retention could affect signaling quantitatively, but is not evidence that this inherited function is absent. Acceptance remains an evolutionary inference; naked-mole-rat immune-output assays have not been located. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Supports the ancestral cGAS signaling function; no target-specific evidence of loss of this function was found. UniProtKB:Q8N884 Β· human CGAS SUPPORTS TRANSFER Supports the ancestral cGAS signaling function; no target-specific evidence of loss of this function was found. MGI:MGI:2442261 Β· mouse Cgas SUPPORTS TRANSFER Supports the ancestral cGAS signaling function; no target-specific evidence of loss of this function was found. Supporting Evidence: file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md Every catalytically important residue that UniProt annotates on human cGAS is present and identical in the naked mole-rat protein PMID:23258413 Overexpression of cGAS activated the transcription factor IRF3 and induced interferon-Ξ² in a STING-dependent manner. file:HETGA/Cgas/Cgas-deep-research-falcon.md Its product is predicted to activate ER-localized STING and downstream TBK1βIRF3 and NF-ΞΊB signaling, promoting type-I interferon and inflammatory gene expression. |
| GO:0002230 positive regulation of defense response to virus by host | IEA GO_REF:0000118 | ACCEPT | Summary: Retain the conserved cGAS contribution to positive regulation of antiviral defense. Reason: The antiviral-defense regulation term concerns inherited cGAMP-STING immune signaling, not the target-specific homologous-recombination phenotype. The curated ancestral cGAS inference is supported by the intact catalytic triad, zinc thumb and nucleotide-binding machinery in A0AAX6RS70 and mammalian cGAMP-STING experiments. PMID:41066557 establishes reversal of the nuclear homologous-recombination phenotype, not loss of innate immune signaling. Prolonged chromatin retention could affect signaling quantitatively, but is not evidence that this inherited function is absent. Acceptance remains an evolutionary inference; naked-mole-rat immune-output assays have not been located. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Supports the ancestral cGAS signaling function; no target-specific evidence of loss of this function was found. Supporting Evidence: file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md Every catalytically important residue that UniProt annotates on human cGAS is present and identical in the naked mole-rat protein PMID:23258413 Overexpression of cGAS activated the transcription factor IRF3 and induced interferon-Ξ² in a STING-dependent manner. |
| GO:0003682 chromatin binding | IEA GO_REF:0000118 | ACCEPT | Summary: Directly and positively confirmed in the naked mole-rat: chromatin binding is the very activity the species-specific adaptation acts on. Reason: This is the one molecular function for which there is explicit naked-mole-rat evidence. The primary study reports that the four species-specific substitutions make cGAS retain chromatin longer after DNA damage, and refers to the resulting state as prolonged chromatin binding. The structural basis is also intact: the arginine anchor by which cGAS docks onto the nucleosome acidic patch (human R255) is conserved as R287 in A0AAX6RS70. Rather than being weakened by the naked-mole-rat divergence, chromatin binding is enhanced by it, so this term is not merely retained but is core. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Node assertion independently confirmed in the target species by PMID:41066557, which is the strongest possible outcome for a propagated term. Supporting Evidence: PMID:41066557 The changes enable cGAS to retain chromatin longer upon DNA damage by weakening TRIM41-mediated ubiquitination and interaction with the segregase P97. PMID:41066557 Prolonged chromatin binding of cGAS enhanced the interaction between repair factors FANCI and RAD50 to facilitate RAD50 recruitment to damage sites, thereby potentiating homologous recombination repair. file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md | nucleosome acidic-patch arginine anchor | R255 | R241 | **R287** | yes | |
| GO:0003690 double-stranded DNA binding | IEA GO_REF:0000118 | ACCEPT | Summary: The dsDNA-reading apparatus, including the zinc thumb, is fully conserved in the naked mole-rat protein. Reason: cGAS reads B-form double-stranded DNA through a zinc thumb and a set of basic DNA-binding surfaces. Residue transfer from human cGAS shows the zinc thumb intact (H390/C396/C397/C404 to H422/C428/C429/C436) and the C-terminal DNA-binding region 384-407 at 62.5% identity, i.e. ordinary orthologue-level conservation with no disruption. The only substitutions among human single-residue annotations that touch DNA are K187 and L195, which UniProt annotates as tuning preference for curved long DNA and which lie in the fast-evolving disordered N-terminal arm; those affect substrate preference at most, not the capacity to bind dsDNA. The native chromatin phenotype is compatible with a DNA-binding protein but, by itself, does not distinguish contacts with DNA from contacts with histones. Nothing about the species-specific C-terminal divergence bears on this. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Supporting Evidence: file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md Every catalytically important residue that UniProt annotates on human cGAS is present and identical in the naked mole-rat protein file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md Global identity to human cGAS is 58.2% over aligned columns (55.2% to mouse) |
| GO:0005634 nucleus | IEA GO_REF:0000118 | ACCEPT | Summary: Supported in the naked mole-rat: the species-specific function is exercised on chromatin after DNA damage, which is nuclear. Reason: The entire naked-mole-rat phenotype is a nuclear one β retention on chromatin after DNA damage, promotion of the FANCI-RAD50 interaction, and RAD50 recruitment to damage sites. A protein cannot do that without a nuclear pool. This is therefore not merely a propagated localization but one corroborated by the species-specific evidence, and it is the compartment in which the core naked-mole-rat activity takes place. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Node assertion corroborated in the target species by PMID:41066557. Supporting Evidence: PMID:41066557 Prolonged chromatin binding of cGAS enhanced the interaction between repair factors FANCI and RAD50 to facilitate RAD50 recruitment to damage sites, thereby potentiating homologous recombination repair. |
| GO:0005829 cytosol | IEA GO_REF:0000118 | ACCEPT | Summary: Canonical cGAS compartment, propagated from a well-grounded node with nothing in the naked-mole-rat data arguing against a cytosolic pool. Reason: Cytosolic localization is the best-established property of cGAS in mammals, and the Euteleostomi node carries it on experimental grounding from both human CGAS and mouse Cgas. Nothing in the naked-mole-rat work excludes a cytosolic pool: what changes in this species is how long the protein stays on chromatin once damage occurs, which shifts the balance between compartments rather than abolishing one of them. The commentary on the primary paper likewise still describes the protein as a cytosolic sensor. A localization claim is a weaker assertion than the functional output that normally accompanies it, and it survives here even though that output is left UNDECIDED β the caveat about the size of the cytosolic pool belongs on the DNA-sensing process terms, not on the existence of the compartment. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Supporting Evidence: PMID:41066590 A DNA repair function in a cytosolic sensor demonstrates a potential role in naked mole-rat longevity. |
| GO:0006974 DNA damage response | IEA GO_REF:0000118 | ACCEPT | Summary: Directly evidenced in the naked mole-rat, where cGAS responds to DNA damage by remaining on chromatin and engaging the repair machinery. Reason: The propagated term is confirmed in the target species. cGAS in the naked mole-rat changes its behaviour specifically upon DNA damage β it is retained on chromatin instead of being extracted β and that change feeds directly into repair-factor recruitment. The term is broad, but here breadth is appropriate: it is the correct parent for both the chromatin-retention step and the downstream recruitment step, and the more specific claim is captured separately by the proposed GO:1905168 row. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Seeded by UniProtKB:Q8N884 alone at this node; the assertion is nevertheless independently confirmed in the naked mole-rat by PMID:41066557. Supporting Evidence: PMID:41066557 The changes enable cGAS to retain chromatin longer upon DNA damage by weakening TRIM41-mediated ubiquitination and interaction with the segregase P97. PMID:41066557 Efficient DNA repair might make possible the longevity of naked mole-rats. |
| GO:0032481 positive regulation of type I interferon production | IEA GO_REF:0000118 | ACCEPT | Summary: Retain the conserved cGAS contribution to positive regulation of type I interferon production. Reason: Type I interferon production is regulated downstream of the conserved cGAMP-STING branch; this is retained as a phylogenetic inference, not asserted as a direct naked-mole-rat cytokine assay. The curated ancestral cGAS inference is supported by the intact catalytic triad, zinc thumb and nucleotide-binding machinery in A0AAX6RS70 and mammalian cGAMP-STING experiments. PMID:41066557 establishes reversal of the nuclear homologous-recombination phenotype, not loss of innate immune signaling. Prolonged chromatin retention could affect signaling quantitatively, but is not evidence that this inherited function is absent. Acceptance remains an evolutionary inference; naked-mole-rat immune-output assays have not been located. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Supports the ancestral cGAS signaling function; no target-specific evidence of loss of this function was found. Supporting Evidence: file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md Every catalytically important residue that UniProt annotates on human cGAS is present and identical in the naked mole-rat protein PMID:23258413 Overexpression of cGAS activated the transcription factor IRF3 and induced interferon-Ξ² in a STING-dependent manner. |
| GO:0035861 site of double-strand break | IEA GO_REF:0000118 | ACCEPT | Summary: Consistent with the naked-mole-rat mechanism, in which cGAS acts on damaged chromatin to drive RAD50 recruitment to damage sites. Reason: The naked-mole-rat abstract does not use the phrase "site of double-strand break", so the direct localization claim is one step beyond what it literally states. But it does say that cGAS is retained on chromatin specifically upon DNA damage and that this facilitates RAD50 recruitment to damage sites β RAD50 being an MRN subunit whose recruitment defines the double-strand break site β and the human orthologue is experimentally documented at double-strand breaks, which is what seeds the node. The species-specific change is to the duration of that residence, not to whether it happens. Accepting is therefore the reading most consistent with the evidence, with the caveat recorded that the naked-mole-rat localization has not been imaged directly. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Seeded at this node by UniProtKB:Q8N884 alone. Donor count is not the issue: the question is whether the naked mole rat still goes to damage sites, and the species-specific evidence indicates it does so for longer, not less. Supporting Evidence: PMID:41066557 Prolonged chromatin binding of cGAS enhanced the interaction between repair factors FANCI and RAD50 to facilitate RAD50 recruitment to damage sites, thereby potentiating homologous recombination repair. |
| GO:0038001 paracrine signaling | IEA GO_REF:0000118 | ACCEPT | Summary: cGAS produces the second messenger used for intercellular antiviral signaling. Reason: GO:0038001 specifies transmission through intercellular fluid. Primary PMID:33768207 establishes export of freely soluble cGAMP and effects on recipient immune cells; this is the relevant paracrine comparator, whereas the gap-junction route in PMID:24077100 alone would not meet that definition. cGAS performs direct work by synthesizing the signal even though other proteins export or import it. Conserved synthesis and the ancestral annotation support transfer to naked-mole-rat cGAS, with species-specific route and output still unmeasured. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS TRANSFER Node grounding is fine; the term is simply a contextual consequence rather than a core activity. Supporting Evidence: PMID:33768207 cGAMP passed through the filter, suggesting that it is exported as a freely soluble molecule file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md Every catalytically important residue that UniProt annotates on human cGAS is present and identical in the naked mole-rat protein |
| GO:0061501 2',3'-cyclic GMP-AMP synthase activity | IEA GO_REF:0000118 | ACCEPT | Summary: The family-defining catalytic activity; every catalytic residue is conserved in the naked-mole-rat protein and the species-specific divergence is elsewhere in the molecule. Reason: This term comes from a different and much deeper node than the rest of the set β PTN000838114 at taxon:6072 (Eumetazoa), seeded by eight members spanning fly cGLRs, zebrafish, mouse and human β and the naked mole rat is well inside it. Since no naked-mole-rat enzymology exists, the propagation was checked against sequence rather than accepted on trust: the catalytic Mg(2+) triad (human E225/D227/D319) transfers intact to E257/D259/D351, the zinc thumb is complete, and the ATP, GTP and cGAMP contacts are conserved. There is no pseudoenzyme signature. Critically, the four residues responsible for the naked mole-rat's altered DNA-repair behaviour all lie in the C-terminal Mab-21-like HhH/H2TH-like domain (437-541), downstream of the zinc thumb and far from the active site, so the functional divergence documented in this species is regulatory and does not touch catalysis. The remaining honest gap is that conserved machinery establishes capability rather than measured activity, which is recorded as a knowledge gap and an experiment rather than by downgrading the call. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN000838114 Β· Eumetazoan mab-21 nucleotidyltransferase node (PTHR10656) SUPPORTS TRANSFER IBD at taxon:6072 with eight seeds across fly, coral, zebrafish, mouse and human; a much deeper placement than the Euteleostomi node carrying the other twelve terms. Supporting Evidence: file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md | catalytic Mg(2+) / 2',3'-cGAMP | D227 | D213 | **D259** | yes | file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md The divergence that reverses the HR phenotype sits in a C-terminal regulatory surface, not in the active site. |
| GO:0071360 cellular response to exogenous dsRNA | IEA GO_REF:0000118 | MODIFY | Summary: Replace the inherited dsRNA-response term with the supported dsDNA-response process. Reason: The focused OpenScientist donor audit found that the four mouse experimental sources behind GO:0071360 test cytosolic DNA, DNA virus, dsDNA binding, or dsDNA-dependent cGAS activation, not exogenous dsRNA or poly(I:C)-driven cGAS signaling. The defect is a dsRNA/dsDNA term-selection error in the donor annotations that then propagated through PTN002579681. Replace the row with cellular response to dsDNA rather than leaving this as an unresolved naked-mole-rat-specific divergence. Propagation Review Root cause: SOURCE BAD Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SOURCE BAD This PAINT source inherits the mouse dsRNA term-selection error even though the donor experiments support DNA-triggered cGAS biology. MGI:MGI:2442261 Β· mouse Cgas SOURCE BAD The four mouse GO:0071360 sources assay dsDNA/cytosolic DNA rather than exogenous dsRNA. Proposed replacements: cellular response to dsDNA Supporting Evidence: file:HETGA/Cgas/Cgas-hypotheses/exogenous-dsrna-response-versus-dna-sensing/openscientist.md none of the four donor papers tests dsRNA PMID:23258413 The cGAS activity was also stimulated by other DNA, including poly(dA:dT), poly(dG:dC) and ISD, but not the RNA poly(I:C). PMID:28363908 binding of nonspecific dsDNA by the N-terminal domain of cGAS promotes its activation. |
| GO:2000042 negative regulation of double-strand break repair via homologous recombination | IEA GO_REF:0000118 | REMOVE | Summary: Contradicted in this species. Naked-mole-rat cGAS does not suppress homologous recombination; it promotes it. This is a sign inversion, not a refinement. Reason: This is the one propagated term with direct, species-specific evidence against it. The primary study states that naked-mole-rat cGAS lacks the suppressive function that the human and mouse orthologs have in homologous recombination repair, and attributes the difference to four amino acids fixed during evolution. The replacement activity runs in the opposite direction: prolonged chromatin retention strengthens the FANCI-RAD50 interaction and promotes RAD50 recruitment to damage sites, potentiating homologous recombination. The propagation is not defective at its source. PTN002579681 sits at taxon:117571 (Euteleostomi) and its seeds β mouse Cgas and human CGAS β genuinely do suppress homologous recombination, so the PAINT curator encoded a real and well-supported property of the species that had been examined. What fails is the assumption that the property is conserved across the whole clade. The naked mole rat is inside that clade and does the opposite, which is exactly the target-specific evidence of functional divergence needed to challenge a node placement β as distinct from an objection based on how many donors seeded it. Retaining a negative-regulation term alongside the positive one would assert both directions at once, so this row is removed rather than modified, and the correct activity is asserted separately as GO:1905168. Propagation Review Root cause: PROPAGATION BAD Failure modes: REGULATORY SIGN INVERSION FUNCTIONAL DIVERGENCE Sources checked: PANTHER:PTN002579681 Β· Euteleostomi cGAS node (PTHR10656) SUPPORTS SOURCE BUT NOT TARGET The IBD is correct for the clade members that were tested; it is the extension to the naked mole rat that fails, because this lineage inverted the sign of the regulation. UniProtKB:Q8N884 Β· human CGAS SUPPORTS SOURCE BUT NOT TARGET Nuclear human cGAS genuinely suppresses homologous recombination; PMID:41066557 uses it as the explicit contrast to the naked-mole-rat protein. MGI:MGI:2442261 Β· mouse Cgas SUPPORTS SOURCE BUT NOT TARGET Same as human; named as a contrasting ortholog in the primary study. Proposed replacements: positive regulation of double-strand break repair via homologous recombination Supporting Evidence: PMID:41066557 naked mole-rat cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) lacks the suppressive function of human or mouse homologs in homologous recombination repair through the alteration of four amino acids during evolution PMID:41066557 Prolonged chromatin binding of cGAS enhanced the interaction between repair factors FANCI and RAD50 to facilitate RAD50 recruitment to damage sites, thereby potentiating homologous recombination repair. file:HETGA/Cgas/Cgas-bioinformatics/RESULTS.md All four positions fall inside the C-terminal Mab-21-like HhH/H2TH-like domain that UniProt annotates at 437-541 on A0AAX6RS70. |
| GO:1905168 positive regulation of double-strand break repair via homologous recombination | IMP PMID:41066557 A cGAS-mediated mechanism in naked mole-rats potentiates DNA... | NEW | Summary: Proposed replacement for GO:2000042. Naked-mole-rat cGAS potentiates homologous recombination repair through prolonged chromatin retention and promotion of RAD50 recruitment. Reason: The primary study reports a positive, mechanistically specified role for cGAS in homologous recombination in this species: prolonged chromatin binding strengthens the FANCI-RAD50 interaction and facilitates RAD50 recruitment to damage sites, thereby potentiating homologous recombination repair. The conclusion is drawn from an engineered four-residue substitution series that shifts the naked-mole-rat protein toward the human and mouse behaviour, which is why IMP is the appropriate evidence code. GO:1905168 was chosen deliberately over the two neighbouring options. The gene product is not itself performing recombination, so GO:0000724 double-strand break repair via homologous recombination would overstate the role; and the effect is specific to the homologous-recombination pathway rather than to repair in general, so the broader GO:2000781 positive regulation of double-strand break repair would understate it. The identifier was confirmed against QuickGO rather than written from memory. No accompanying ageing or lifespan term is proposed. The abstract does support an anti-ageing role, but reduced senescence and extended lifespan here are indirect, downstream consequences of improved repair, and NEW is explicitly not for indirect or pleiotropic effects. Supporting Evidence: PMID:41066557 Prolonged chromatin binding of cGAS enhanced the interaction between repair factors FANCI and RAD50 to facilitate RAD50 recruitment to damage sites, thereby potentiating homologous recombination repair. PMID:41066557 naked mole-rat cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) lacks the suppressive function of human or mouse homologs in homologous recombination repair through the alteration of four amino acids during evolution |
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Download this section (compressed HTML)Q: How do naked-mole-rat cGAMP production and STING/interferon dose responses compare with those of human and mouse cGAS? The inherited signaling annotation is supported, but its quantitative regulation remains unmeasured.
Suggested experts: Zhiyong Mao, Yu Chen
Q: Do the four C-terminal substitutions affect only TRIM41 and p97 engagement, or do they also change cGAS turnover, oligomerisation or nucleosome affinity in ways that would alter the enzyme's regulation more broadly?
Suggested experts: Zhiyong Mao
Q: Given that the chromatin retention which potentiates repair corresponds, in human and mouse, to the catalytically restrained nucleosome-bound state, is there a trade-off in the naked mole rat between genome maintenance and cytosolic DNA surveillance?
Suggested experts: Vera Gorbunova, Andrei Seluanov
Q: Is the homologous-recombination-promoting activity of naked-mole-rat cGAS independent of its nucleotidyltransferase activity, as would be expected if it acts as a chromatin-bound scaffold rather than through cGAMP?
Suggested experts: Zhiyong Mao
Q: Should mouse Cgas GO:0071360 annotations and PANTHER PTN002579681 propagation be corrected to use GO:1990786 cellular response to dsDNA rather than a dsRNA-response term?
Experiment: Express and purify recombinant A0AAX6RS70 alongside human CGAS as a positive control, and measure 2'3'-cGAMP production from ATP and GTP in the presence and absence of activating double-stranded DNA by LC-MS/MS. Include a triad mutant (E257A/D259A) as a negative control. This would convert the sequence-level inference behind the GO:0061501 call into a direct measurement.
Hypothesis: Naked-mole-rat cGAS retains 2'3'-cGAMP synthase activity, as predicted by the complete conservation of its catalytic triad, zinc thumb and nucleotide contacts.
Type: in vitro enzyme assay
Experiment: Transfect naked-mole-rat and mouse fibroblasts with cytosolic double-stranded DNA and with poly(I:C), and compare cGAMP levels, STING and IRF3 phosphorylation, and interferon-beta and interferon-stimulated gene induction, with cGAS knockdown as the specificity control. A positive result would strengthen the accepted GO:0002218, GO:0002230 and GO:0032481 inferences; a negative result would be the first evidence that the immune arm has diverged in this lineage.
Hypothesis: The cGAS-STING-interferon axis is functional in naked-mole-rat cells, so the four propagated innate-immune annotations transfer despite the chromatin-retention adaptation.
Type: cellular innate immune signalling assay
Experiment: Induce site-specific double-strand breaks and follow endogenous cGAS by live-cell imaging and by chromatin immunoprecipitation at the break site in naked-mole-rat cells, comparing residence times with mouse cells. This would directly test the GO:0035861 localization, which is currently accepted on mechanistic inference rather than on imaging in this species.
Hypothesis: Naked-mole-rat cGAS localises to double-strand break sites, as the propagated cellular component term asserts and as its role in RAD50 recruitment implies.
Type: laser microirradiation and ChIP at induced breaks
Experiment: Compare homologous recombination reporter efficiency in cells complemented with wild-type naked-mole-rat cGAS, a catalytically dead triad mutant, and the humanising four-residue variant. Separating catalysis from the repair role would determine whether the two core functions recorded here are genuinely independent activities of the same protein.
Hypothesis: The promotion of homologous recombination by naked-mole-rat cGAS is a scaffolding function that does not require nucleotidyltransferase activity.
Type: DR-GFP homologous recombination reporter assay
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