id: A0A6I8TLE4 gene_symbol: A0A6I8TLE4 product_type: PROTEIN status: DRAFT taxon: id: NCBITaxon:7159 label: Aedes aegypti description: >- A0A6I8TLE4 is the 1442-residue AAEL019982-PE splice product of Aedes aegypti gene AAEL019982 (NCBI Gene 5579847), one of five UniProt products of this gene that differ only in their first ~130 residues. It is the mosquito ortholog of Drosophila raskol, sharing PANTHER subfamily PTHR10194:SF60 and 61% sequence identity with it, and belongs to the DAB2IP/RASAL2/SynGAP-like branch of the Ras GTPase-activating protein family. The protein has an N-terminal SynGAP-like PH domain, a C2 domain, a central RasGAP domain that retains the catalytic arginine finger (R483, in a TFRGNS finger loop identical to raskol and neurofibromin), a long disordered middle region, and a C-terminal DAB2IP-type (DAB2P_C) region. By homology it is expected to accelerate GTP hydrolysis by a Ras-subfamily small GTPase and so to damp Ras-type signalling; its substrate has not been identified. The Drosophila ortholog localises to the plasma membrane and apical cell cortex of epithelial cells, colocalises with E-cadherin at cell-cell contacts and is needed for border-cell adhesion and actin-protrusion control. The mosquito protein itself has not been studied experimentally. references: - id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-deep-research-falcon.md title: Deep research report for A0A6I8TLE4 (falcon) reference_review: relevance: LOW correctness: LOW_QUALITY review_notes: >- Re-review 2026-10-21. The report states that no contexts were retrieved, so it is an ungrounded model answer. It correctly says the protein is uncharacterised, but it misreads IPR039360 (the InterPro entry for the PANTHER RasGAP family) as a Ras GTPase switch domain, invokes intersectins as architectural relatives, and never identifies the protein as a RasGAP or as the raskol ortholog. No longer used as support for any function claim; kept for provenance. - id: GO_REF:0000002 title: Gene Ontology annotation through association of InterPro records with GO terms reference_review: relevance: LOW correctness: VERIFIED review_notes: >- Re-review 2026-10-21. Previously used as the original_reference_id of the proposed annotations, although none of them derives from InterPro2GO and GOA holds no rows for this accession. No longer cited by any annotation; kept for provenance. - id: GO_REF:0000104 title: Electronic Gene Ontology annotations created by UniProt reference_review: relevance: LOW correctness: VERIFIED review_notes: >- Re-review 2026-10-21. Not cited by any annotation; kept for provenance. - id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-bioinformatics/RESULTS-rereview.md title: A0A6I8TLE4 re-review bioinformatics (subfamily comparison, arginine finger, isoforms) reference_review: relevance: HIGH correctness: VERIFIED review_notes: >- Reproducible MAFFT comparisons written for this re-review (compare_rasgap_rereview.py, compare_isoforms_rereview.py) with their raw outputs saved next to the results file. - id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-uniprot.txt title: UniProt record A0A6I8TLE4 - id: PMID:30763317 title: Evolutionary rate covariation analysis of E-cadherin identifies Raskol as a regulator of cell adhesion and actin dynamics in Drosophila. reference_review: relevance: HIGH correctness: VERIFIED review_notes: >- Full text cached. Only experimental study of the Drosophila ortholog raskol (CG42684, Q8T498): plasma-membrane and apical localisation, colocalisation with DE-cadherin, RNAi phenotypes in border-cell migration. GAP activity is not measured; the authors infer it from the domain. - id: PMID:9619631 title: Characterization of the C. elegans gap-2 gene encoding a novel Ras-GTPase activating protein and its possible role in larval development. reference_review: relevance: MEDIUM correctness: VERIFIED review_notes: >- Abstract only. gap-2 (Q8MLZ5) is an SF60 member; isolated by complementing a fission yeast gap1 mutant, and genetically a negative regulator of LET-60 Ras. Supports Ras-directed GAP function in the subfamily. - id: PMID:8371782 title: BUD2 encodes a GTPase-activating protein for Bud1/Rsr1 necessary for proper bud-site selection in yeast. reference_review: relevance: MEDIUM correctness: VERIFIED review_notes: >- Abstract only. Purified BUD2 (P33314, SF60) stimulates GTP hydrolysis by the Ras-like GTPase Bud1/Rsr1. Direct biochemical evidence of GAP activity in the subfamily; the substrate is a fungal Rap-like GTPase and is not transferred to the mosquito protein. existing_annotations: - term: id: GO:0005096 label: GTPase activator activity qualifier: enables evidence_type: ISS original_reference_id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-bioinformatics/RESULTS-rereview.md review: summary: >- Proposed annotation (GOA holds no rows for this accession). The protein has a complete RasGAP domain (IPR001936, 391-735) with the conserved site and retains the catalytic arginine finger (R483, TFRGNS). GTPase activator activity is the IBD assertion PAINT placed at the PTHR10194 root node PTN008939999, from which A0A6I8TLE4 descends, and within its own subfamily SF60 purified BUD2 stimulates GTP hydrolysis by its Ras-like target and gap-2 complements a fission yeast RasGAP mutant. action: NEW reason: >- Family-wide catalytic function with an intact catalytic interface in the target. Kept at the general GTPase activator level because the substrate GTPase is not known for this protein or for raskol, and SF60 members target different Ras-subfamily GTPases. supported_by: - reference_id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-bioinformatics/RESULTS-rereview.md supporting_text: >- The catalytic arginine finger of the RasGAP domain is conserved in A0A6I8TLE4. - reference_id: PMID:8371782 supporting_text: >- Bud2p purified from yeast stimulates GTP hydrolysis by Bud1p - reference_id: PMID:9619631 supporting_text: C. elegans gap-2 encodes a novel Ras-GAP - term: id: GO:1902531 label: regulation of intracellular signal transduction qualifier: involved_in evidence_type: ISS original_reference_id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-bioinformatics/RESULTS-rereview.md review: summary: >- Proposed annotation replacing the earlier proposals of GO:0007265 Ras protein signal transduction and GO:0046580 negative regulation of Ras protein signal transduction. This is the process term PAINT placed at the PTHR10194 root node alongside GTPase activator activity, and the term raskol, gap-2 and BUD2 carry by IBA. A GAP acts on the GTPase of a signalling cassette rather than relaying the signal, so GO:0007265 was the wrong relation; and although one of the node's seeds, human DAB2IP (Q5VWQ8), has an experimental (IMP) annotation to GO:0046580, PAINT curators propagated only the general regulation term, which is treated here as a deliberate decision given the varied substrates within the family. action: NEW reason: >- Follows the PAINT node placement and the conserved catalytic domain. A more specific Ras-pathway term would need evidence on the substrate GTPase for this branch. supported_by: - reference_id: PMID:9619631 supporting_text: >- GAP-2 appears to function as a negative regulator of LET-60 Ras during larval development - reference_id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-bioinformatics/RESULTS-rereview.md supporting_text: >- Drosophila raskol is by far the closest relative and shares the PANTHER subfamily, so it is the dipteran ortholog. - term: id: GO:0005886 label: plasma membrane qualifier: located_in evidence_type: ISS original_reference_id: PMID:30763317 review: summary: >- Proposed annotation by transfer from the Drosophila ortholog raskol (same PANTHER subfamily SF60, 61% identity), which localises to the cell membrane and apical surface of epithelial cells and colocalises with DE-cadherin at cell contacts. The target has the same SynGAP-like PH and C2 membrane-binding modules. The earlier proposal of cytosol was withdrawn: no evidence placed the ortholog in the cytosol rather than at the membrane and cortex. action: NEW reason: >- Ortholog localisation, consistent with the conserved PH and C2 domains. Placed outside core function because it rests on a single ortholog study. supported_by: - reference_id: PMID:30763317 supporting_text: >- In Drosophila stage 8 embryos, before BCs have delaminated, Raskol localized to the cell membrane of BCs and PCs and was enriched at the PC apical membrane core_functions: - description: >- Ras-family GTPase-activating protein of the raskol/DAB2IP-like branch. Its RasGAP domain, with a conserved catalytic arginine finger, is expected to accelerate GTP hydrolysis by a Ras-subfamily small GTPase and thereby limit Ras-type intracellular signalling; the substrate GTPase has not been identified. molecular_function: id: GO:0005096 label: GTPase activator activity directly_involved_in: - id: GO:1902531 label: regulation of intracellular signal transduction locations: - id: GO:0005886 label: plasma membrane supported_by: - reference_id: file:AEDAE/A0A6I8TLE4/A0A6I8TLE4-bioinformatics/RESULTS-rereview.md supporting_text: >- The catalytic arginine finger of the RasGAP domain is conserved in A0A6I8TLE4. - reference_id: PMID:8371782 supporting_text: >- Bud2p purified from yeast stimulates GTP hydrolysis by Bud1p - reference_id: PMID:30763317 supporting_text: >- In Drosophila stage 8 embryos, before BCs have delaminated, Raskol localized to the cell membrane of BCs and PCs and was enriched at the PC apical membrane suggested_questions: - question: >- Which small GTPase does the raskol branch act on? Human DAB2IP and RASAL2 act on Ras and SynGAP on Ras and Rap, yeast BUD2 on the Rap-like Rsr1, and C. elegans gap-2 genetically on LET-60 Ras. If insect raskol is a Ras GAP, GO:0046580 negative regulation of Ras protein signal transduction would be justified for the branch; PAINT currently assigns only GO:1902531 at the family root. - question: >- Do the five AAEL019982 products, which differ only in their first ~130 residues (partly within the broad PH-domain region), differ in membrane targeting or tissue expression? - question: >- Does the mosquito protein, like Drosophila raskol, colocalise with E-cadherin at adherens junctions? suggested_experiments: - hypothesis: >- The RasGAP domain of A0A6I8TLE4 stimulates GTP hydrolysis by a mosquito Ras-subfamily GTPase, and the R483A arginine-finger mutation abolishes it. description: >- Purify the RasGAP domain (wild type and R483A) and measure single-turnover GTP hydrolysis by recombinant A. aegypti Ras, Rap1 and R-Ras homologs (phosphate-release or fluorescent-nucleotide assays). experiment_type: biochemical assay - hypothesis: >- A0A6I8TLE4 localises to the plasma membrane and cell contacts through its PH and C2 domains. description: >- Express tagged full-length protein and PH/C2 deletion variants in Aag2 cells and score cortical versus cytosolic localisation and colocalisation with E-cadherin. experiment_type: cell biology / microscopy