FANCE evidence and source guide

This agent-authored guide combines the original-evidence and database-source assessments. Their separate complete reading scopes remain labeled in the reference notes; an image read by one assessment is not attributed to the other.

FANCE is a nuclear FA core component that recognizes the FANCD2 substrate. Association with another subunit, substrate recognition and ubiquitin-transfer chemistry are different claims. The sources below identify which experiments establish each claim. Natural alternative products are not asserted: the official displayed protein has 536 residues, while tagged proteins, engineered deletions and external transcript models have their own boundaries.

Substrate recognition and core function

PMID:12093742, Results and Methods, Figures 1, 4 and 6 reports endogenous and transfected human FANCE in HeLa nuclear foci, association with the FA core, mammalian two-hybrid interactions, and FANCE–FANCC/FANCD2 preparations from insect cells. Figure 1 and Figure 6 pixels were inspected in the direct-original assessment; the independent database assessment read the stated Methods, Results and captions without inspecting those pixels. Figure 6 supports FANCE–FANCD2 binding through complementary assays; it is not a purified ubiquitin-transfer experiment or a simultaneous stoichiometry determination of the entire substrate-bound complex. The discussion expressly leaves FANCC cotransport versus nuclear retention unresolved.

PMID:24451376, Methods and Results, Figures 1–2 provides a more specific functional distinction. Human FANCE CΔ25/F522D perturbations impair FANCD2 recognition and downstream monoubiquitination while retaining measured associations with other core subunits. Human complementation, cross-linker sensitivity and chromosome endpoints connect this recognition step to repair. Figures 1–2 pixels were inspected in the direct-original assessment; the independent database assessment read the stated Methods, Results and Discussion without inspecting those pixels. The authors assume retained intrinsic catalytic competence of the mutant complex and cannot exclude additional affected interactions; those limits remain part of the interpretation. No autonomous ubiquitin-transfer activity is assigned to FANCE.

The inherited proposal's original reference is PMID:12093742; PMID:12649160 is a supporting reference. Its authored proposal is refined in place, preserving its original PMID, evidence and five quotation anchors. The specific term, GO:1990756 ubiquitin-like ligase-substrate adaptor activity, describes bringing a ubiquitin-like ligase and its substrate together. It is supported here by the recognition evidence rather than generic membership in a stable complex. The current GO:0030674 comment distinguishes stable hold-together scaffolds, for which it points to GO:0140378. FANCL/UBE2T, rather than FANCE, perform ubiquitin transfer.

Current official human comparator queries recover GO:1990756 for VHL (IDA PMID:24899725/34290272) and FBXW7 (IDA PMID:15150404/34741373, including an isoform-specific record). The bounded SPOP query has no exact or descendant hit. This mixed result does not establish a universal convention or a missing-annotation claim. The fixed-source GO-CAM index has no FANCE/Q9HB96 entry. The specific authored proposal now occupies its original position with action NEW; the earlier broad term and duplicate last proposal are no longer asserted. This is an in-place refinement of an authored annotation, not a change to an imported GOA term, and it adds no distinct activity or quotation. One core unit represents this substrate-recognition activity in interstrand cross-link repair, with nucleoplasm as a supported functional pool and FA nuclear-complex membership recorded separately. The retained broad nucleus contexts allow other nuclear pools. The chromatin annotation is accepted by component-level projection within the chromatin-associated FA repair complex, while the core retains its supported nucleoplasm location.

Nuclear and chromatin evidence

PMID:11001585(https://pmc.ncbi.nlm.nih.gov/articles/PMC1288571/) establishes the human FANCE cDNA by complementation of MMC hypersensitivity. Its nuclear-localization signals are predictions, not imaging. Independent direct nuclear imaging in PMID:12093742 supports the broad nucleus annotation. Nuclear foci do not by themselves establish an exclusively chromatin-bound pool.

PMID:22343915, main Figures 1 and 5 identifies FANCE in affinity-purified human HeLaS3 nuclear core material. Main Figure 1A/B explicitly labels FANCE; these pixels were inspected. The chromatin experiment in Figure 5H instead measures FANCA, FANCG, FAAP20 and H2A. The authentic Blood supplemental PDF supplies FANCE construct and fractionation context; Figure S1C (PDF page 11, pixels inspected) labels other core components but not FANCE. The imported chromatin IDA is accepted as a legitimate component-level projection within this chromatin-associated repair complex. Independent human FANCE core-membership and FANCD2-recruitment experiments corroborate its role in that machinery; no separately labeled FANCE chromatin-fraction measurement is invented. The figure-level limitation does not establish absence, and the accepted association does not make every nuclear FANCE molecule chromatin-bound.

PMID:20347428, Results and Figure 1B/C, explicitly lists FANC-E in FANCM immunoprecipitates from fractionated human HeLa nuclear extract, with mass spectrometry/immunoblotting and gel-filtration/IP Methods. The separate small FANCM–MHF complex and its DNA-remodeling assays do not establish those activities for FANCE. The complete BioC main was available and selected operative text was read; a later direct HTML route was a non-article page and no new pixels are claimed.

PMID:22266823, Results and Figure 2a/e identifies FANCE with FANCA/FANCC in FLAG-FAAP20 immunoprecipitates from human 293T cells and separately measures FANCE abundance after FAAP20 depletion/MG132 in HeLa cells. The in-vitro direct-binding experiment in Figure 2b tests FANCA–FAAP20. The online Methods are referenced by the available main but were not separately read; no supplement or pixels were inspected for this paper.

Partner-specific binding contexts

The PMID:12649160 abstract positively reports FANCC–FANCE mapping and disease-mutant effects. Its operative full text was not recovered; failed original routes are retained in the evidence record. The same partner association is independently supported by the actual PMID:12093742 experiments and BioPlex. This supports retaining the generic binding context as non-core without pretending to have verified the old constructs or assays. The FANCC-facing interaction alone does not establish a transport mechanism or the separately assessed FANCD2 substrate-recognition function.

For PMID:33961781, actual official BioPlex tables identify FANCC/Q00597 bait and FANCE/Q9HB96 prey in HCT116: BioPlex_HCT116_Network_5.5K_Dec_2019.tsv, line 32947; BioPlex_3.0_HCT116_DirectedEdges.tsv, line 38136. The corresponding inspected 293T release tables do not contain the pair, which is not biological nonbinding. Actual anti-HA AP-MS/cell-line Methods were read. These data establish cellular coassociation, not necessarily a purified binary interface, reciprocal recovery or equivalence of every current protein model. Supported generic binding is retained as non-core under the standing campaign exception.

For PMID:35512704(https://pmc.ncbi.nlm.nih.gov/articles/PMC9597701/), actual Data S1, PDF page 3, Figure iiiA, FANCE panel, shows tagged FANCE recovered with GST-AKT1 WT and E17K in HEK293T; the pixels were inspected without densitometry. Table S3, sheet S3B_AKT1-E17K Go-PPI evaluation, row 22, records positive GST pull-down/HEK293T and NanoPCA/MCF7 for AKT1-E17K–FANCE, with semi-endogenous and endogenous co-IP marked N/A. Figure iiiB does not test FANCE. The Table S2 download was not a valid complete XLSX and its quantitative matrix is not claimed read. GO:0043422 protein kinase B binding names the observed partner class; it does not assign kinase regulation, FANCE phosphorylation or an endogenous repair mechanism. This remains a contextual interaction outside the cross-link-repair core despite the more informative partner-specific term.

Repair pathway and source boundaries

PMID:19965384 directly tests replication-coupled repair of a defined cisplatin interstrand cross-link in Xenopus extracts using FANCI/FANCD2 depletion and reconstitution. It is pathway context for the imported FANCE NAS. Human FANCE recognition/complementation evidence comes from PMID:12093742 and PMID:24451376. FANCE contributes to substrate recruitment; incision, lesion-bypass synthesis and FANCL/UBE2T ubiquitin-transfer chemistry are not transferred to it by pathway membership.

Reference assessments distinguish three things: immutable normal-cache metadata, external original availability, and the passages or images actually read. The full_text_unavailable flags follow the selected normal-cache convention used in this campaign; the accompanying assessments explicitly identify complete external originals and bounded reading. Failed BioC/HTML/PDF and workbook routes are retained, not silently relabeled as originals. Inherited quotation text and reference associations remain unchanged; no new publication quotation is added. This authored source guide records evidence and locators and is not a generated provider research file.

Human Protein Atlas and nuclear breadth

The current FANCE Human Protein Atlas subcellular page and official XML report supported nucleoplasm localization with two antibodies. HPA013343 covers HEL, THP-1 and U2OS; CAB014893 covers A-431, U-251MG and U2OS. Actual HEL, A-431 and THP-1 merged images were inspected. Green signal lies in nuclear interiors, with excluded nucleolar regions especially clear in A-431. No quantitative image analysis was performed. The downloaded U2OS image was not independently viewed.

HPA013343's 141-residue antigen maps exactly to displayed Q9HB96 residues 112–252. CAB014893 has no antigen sequence in the inspected record. The data also report mitotic chromosome and centrosome in THP-1, so nucleoplasm is not presented as the sole cellular location. The HPA013343 record retains release version 22; a current retrieval date is not a new experiment. The broad nucleus term is retained alongside source-specific nucleoplasm, whose definition excludes chromosomes. Neither record establishes the localization of every external RefSeq/Ensembl protein model.

Family and phylogenetic evidence

The current InterPro IPR039685 FANCE family matches human Q9HB96 and maps to interstrand cross-link repair and Fanconi anaemia nuclear-complex membership. The mapping is corroborated by the human experiments above.

Actual PANTHER PTHR32094 version 19 tree and alignment place the 536-residue human target PTN002528904 beneath PTN001280497. The target alignment ungaps exactly to displayed Q9HB96. The actual IBD assertion for GO:0043240 at that ancestor uses human Q9HB96 experimental evidence; no NOT was found on the target path. This self-WITH is legitimate descendant grounding, not circularity. The tree, alignment and source assertion support the inherited complex-membership annotation; they do not establish a catalytic role.

Reactome target compartments

All eleven assigned events were traced to FANCE's own physical entity, rather than borrowing the compartment of an unrelated participant. Human Q9HB96 residues 1–536 occur as FANCE in nucleoplasm, entity 419543, or as FANCE in cytosol, entity 420786. The ten nuclear events retain a modeled nucleoplasmic pool; they are not ten independent imaging experiments.

The PKR event emphasizes FANCC binding and FANCA/FANCC/FANCG cellular evidence. Its explicit FANCE component supports the modeled location but does not establish a FANCE-specific PKR mechanism. Event summations and literature identifiers were inspected; not every cited original was read. All 26 target routes are complete in the saved graph; eight unexpanded off-target objects remain a limitation of that graph capture.

Nuclear-periphery transfer: measured protein differs from the donor assignment

The extra current human GO:0034399 IEA comes from Ensembl-Compara transfer of mouse Fance B8JJD3/ENSMUSP00000110451. Its current MGI IDA donor is Fu et al., 2016 (PMID:27486799). The complete official original was recovered. Mouse strain/mutation and RNA Methods, immunohistochemistry Methods, Table 1, Results “Fancd2,” Figure 8 caption and pixels, and relevant Discussion were inspected.

The measured perinuclear antigen is Fancd2, detected with antibody NB100-182. The authors explicitly state that their Fance immunohistology attempts were unsuccessful. Wild-type germ cells show Fancd2 around the nuclear membrane, whereas Fance-mutant testes do not show that pattern. This supports a Fance-dependent effect on another protein's localization; it does not locate Fance itself. The electronic transfer is therefore proposed for removal, with a precise, unreported MGI query about target attribution. No corrected gene accession is guessed. This is not evidence that human FANCE cannot localize to the nuclear periphery and does not justify a NOT annotation.

Protein and evidence limits

The official displayed Q9HB96 sequence has 536 residues, with no alternative-products comment. Its full-length chain feature is not evidence for a processed shorter product. Two RefSeq NP records, sixteen XP records and unresolved Ensembl lookups remain external model information; no measured localization or activity is transferred to an untested model. Tagged full-length FANCE, the engineered 273–536 fragment, C-terminal deletions and Phe522 substitutions retain their stated assay identities.

The human recognition studies do not establish the complete geometry of a substrate-loaded FA core, prove that every mutant preserves intrinsic catalytic competence, or distinguish FANCC import from retention. These questions motivate controlled reconstitution and endogenous localization experiments. They do not require additional process annotations or a second molecular-function unit.

All 23 inherited quotation paths and reference associations are retained. The original authored position 25 now proposes GO:1990756 with action NEW and keeps its five quotation anchors, original PMID and evidence. The duplicate last proposal at 27 is removed; the genuine current nuclear-periphery context at 26 remains unchanged and reviewed as REMOVE. The imported sources and seven source-derived supporting-entity lists remain exact. This yields 27 active objects: 26 imported contexts and one specific authored proposal (21 ACCEPT, 3 KEEP_AS_NON_CORE, 1 MODIFY, 1 REMOVE, 1 NEW). The single core and all 31 reference assessments remain unchanged. The historical journal records the prior representation. No upstream report has been submitted, and neither a missing staining signal nor an unavailable original is used as a NOT annotation.

CHEK1 phosphorylation and cross-link resistance

PMID:17296736, original Methods, Results, Figures 1–2 and Table 1 establishes CHEK1-mediated phosphorylation of human FANCE at Thr346 and Ser374. Full-length FLAG-tagged human FANCE and the TS/AA mutant were expressed in patient-derived FA-E cells; bacterial GST fragments spanning residues 332–365, 349–382 or 332–382 and purified human-FANCE residues 149–536 were used in recombinant kinase assays. The kinase is Chk1, not FANCE. Phosphosite-mutant and kinase controls support the site assignment; the mutant restores basal FANCD2 monoubiquitination and nuclear foci but does not restore normal damage-induced increases or mitomycin C resistance. These distinct outcomes should not be compressed into a claim that all FANCD2 regulation is phosphorylation-independent. The downstream phosphorylation-dependent mechanism remains unresolved.

For this clarification, the complete official PMC main HTML was recovered and the stated Methods, Results, figure captions, Table 1 and relevant Discussion were read. No new figure pixels or supplements were inspected. The selected immutable normal cache is labeled full-text-available but contains the abstract and Discussion; that metadata and its bytes remain unchanged. The newly read Methods/Results are sourced to the separate official original, not retroactively attributed to the earlier cache or reading record.