RHGF_DROME
ID RHGF_DROME Reviewed; 1025 AA.
AC X2JDY8; Q86NY4; Q9VXQ2;
DT 29-SEP-2021, integrated into UniProtKB/Swiss-Prot.
DT 11-JUN-2014, sequence version 1.
DT 03-AUG-2022, entry version 69.
DE RecName: Full=Rho GTPase-activating protein Graf {ECO:0000305};
DE AltName: Full=GTPase regulator associated with focal adhesion kinase {ECO:0000312|FlyBase:FBgn0030685};
GN Name=Graf {ECO:0000312|FlyBase:FBgn0030685};
GN ORFNames=CG8948 {ECO:0000312|FlyBase:FBgn0030685};
OS Drosophila melanogaster (Fruit fly).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Endopterygota; Diptera; Brachycera; Muscomorpha; Ephydroidea;
OC Drosophilidae; Drosophila; Sophophora.
OX NCBI_TaxID=7227 {ECO:0000312|Proteomes:UP000000803};
RN [1] {ECO:0000312|Proteomes:UP000000803}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=Berkeley {ECO:0000312|Proteomes:UP000000803};
RX PubMed=10731132; DOI=10.1126/science.287.5461.2185;
RA Adams M.D., Celniker S.E., Holt R.A., Evans C.A., Gocayne J.D.,
RA Amanatides P.G., Scherer S.E., Li P.W., Hoskins R.A., Galle R.F.,
RA George R.A., Lewis S.E., Richards S., Ashburner M., Henderson S.N.,
RA Sutton G.G., Wortman J.R., Yandell M.D., Zhang Q., Chen L.X., Brandon R.C.,
RA Rogers Y.-H.C., Blazej R.G., Champe M., Pfeiffer B.D., Wan K.H., Doyle C.,
RA Baxter E.G., Helt G., Nelson C.R., Miklos G.L.G., Abril J.F., Agbayani A.,
RA An H.-J., Andrews-Pfannkoch C., Baldwin D., Ballew R.M., Basu A.,
RA Baxendale J., Bayraktaroglu L., Beasley E.M., Beeson K.Y., Benos P.V.,
RA Berman B.P., Bhandari D., Bolshakov S., Borkova D., Botchan M.R., Bouck J.,
RA Brokstein P., Brottier P., Burtis K.C., Busam D.A., Butler H., Cadieu E.,
RA Center A., Chandra I., Cherry J.M., Cawley S., Dahlke C., Davenport L.B.,
RA Davies P., de Pablos B., Delcher A., Deng Z., Mays A.D., Dew I.,
RA Dietz S.M., Dodson K., Doup L.E., Downes M., Dugan-Rocha S., Dunkov B.C.,
RA Dunn P., Durbin K.J., Evangelista C.C., Ferraz C., Ferriera S.,
RA Fleischmann W., Fosler C., Gabrielian A.E., Garg N.S., Gelbart W.M.,
RA Glasser K., Glodek A., Gong F., Gorrell J.H., Gu Z., Guan P., Harris M.,
RA Harris N.L., Harvey D.A., Heiman T.J., Hernandez J.R., Houck J., Hostin D.,
RA Houston K.A., Howland T.J., Wei M.-H., Ibegwam C., Jalali M., Kalush F.,
RA Karpen G.H., Ke Z., Kennison J.A., Ketchum K.A., Kimmel B.E., Kodira C.D.,
RA Kraft C.L., Kravitz S., Kulp D., Lai Z., Lasko P., Lei Y., Levitsky A.A.,
RA Li J.H., Li Z., Liang Y., Lin X., Liu X., Mattei B., McIntosh T.C.,
RA McLeod M.P., McPherson D., Merkulov G., Milshina N.V., Mobarry C.,
RA Morris J., Moshrefi A., Mount S.M., Moy M., Murphy B., Murphy L.,
RA Muzny D.M., Nelson D.L., Nelson D.R., Nelson K.A., Nixon K., Nusskern D.R.,
RA Pacleb J.M., Palazzolo M., Pittman G.S., Pan S., Pollard J., Puri V.,
RA Reese M.G., Reinert K., Remington K., Saunders R.D.C., Scheeler F.,
RA Shen H., Shue B.C., Siden-Kiamos I., Simpson M., Skupski M.P., Smith T.J.,
RA Spier E., Spradling A.C., Stapleton M., Strong R., Sun E., Svirskas R.,
RA Tector C., Turner R., Venter E., Wang A.H., Wang X., Wang Z.-Y.,
RA Wassarman D.A., Weinstock G.M., Weissenbach J., Williams S.M., Woodage T.,
RA Worley K.C., Wu D., Yang S., Yao Q.A., Ye J., Yeh R.-F., Zaveri J.S.,
RA Zhan M., Zhang G., Zhao Q., Zheng L., Zheng X.H., Zhong F.N., Zhong W.,
RA Zhou X., Zhu S.C., Zhu X., Smith H.O., Gibbs R.A., Myers E.W., Rubin G.M.,
RA Venter J.C.;
RT "The genome sequence of Drosophila melanogaster.";
RL Science 287:2185-2195(2000).
RN [2] {ECO:0000312|Proteomes:UP000000803}
RP GENOME REANNOTATION.
RC STRAIN=Berkeley {ECO:0000312|Proteomes:UP000000803};
RX PubMed=12537572; DOI=10.1186/gb-2002-3-12-research0083;
RA Misra S., Crosby M.A., Mungall C.J., Matthews B.B., Campbell K.S.,
RA Hradecky P., Huang Y., Kaminker J.S., Millburn G.H., Prochnik S.E.,
RA Smith C.D., Tupy J.L., Whitfield E.J., Bayraktaroglu L., Berman B.P.,
RA Bettencourt B.R., Celniker S.E., de Grey A.D.N.J., Drysdale R.A.,
RA Harris N.L., Richter J., Russo S., Schroeder A.J., Shu S.Q., Stapleton M.,
RA Yamada C., Ashburner M., Gelbart W.M., Rubin G.M., Lewis S.E.;
RT "Annotation of the Drosophila melanogaster euchromatic genome: a systematic
RT review.";
RL Genome Biol. 3:RESEARCH0083.1-RESEARCH0083.22(2002).
RN [3] {ECO:0000312|EMBL:AAO39581.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM A).
RC STRAIN=Berkeley {ECO:0000312|EMBL:AAO39581.1};
RC TISSUE=Embryo {ECO:0000312|EMBL:AAO39581.1};
RA Stapleton M., Brokstein P., Hong L., Agbayani A., Carlson J., Champe M.,
RA Chavez C., Dorsett V., Dresnek D., Farfan D., Frise E., George R.,
RA Gonzalez M., Guarin H., Kronmiller B., Li P., Liao G., Miranda A.,
RA Mungall C.J., Nunoo J., Pacleb J., Paragas V., Park S., Patel S.,
RA Phouanenavong S., Wan K., Yu C., Lewis S.E., Rubin G.M., Celniker S.;
RL Submitted (FEB-2003) to the EMBL/GenBank/DDBJ databases.
RN [4] {ECO:0000305}
RP FUNCTION, INTERACTION WITH EGFR, SUBCELLULAR LOCATION, DEVELOPMENTAL STAGE,
RP AND DISRUPTION PHENOTYPE.
RX PubMed=28993397; DOI=10.1242/dev.153288;
RA Kim S., Nahm M., Kim N., Kwon Y., Kim J., Choi S., Choi E.Y., Shim J.,
RA Lee C., Lee S.;
RT "Graf regulates hematopoiesis through GEEC endocytosis of EGFR.";
RL Development 144:4159-4172(2017).
RN [5] {ECO:0000305}
RP FUNCTION, SUBCELLULAR LOCATION, DOMAIN, DISRUPTION PHENOTYPE, AND
RP MUTAGENESIS OF 28-GLU--HIS-1025 AND 393-PRO--ILE-588.
RX PubMed=33835025; DOI=10.7554/elife.63535;
RA Sharma S., Rikhy R.;
RT "Spatiotemporal recruitment of RhoGTPase protein GRAF inhibits actomyosin
RT ring constriction in Drosophila cellularization.";
RL Elife 10:0-0(2021).
RN [6] {ECO:0000305}
RP FUNCTION, TISSUE SPECIFICITY, AND DISRUPTION PHENOTYPE.
RX PubMed=33892766; DOI=10.1186/s13041-021-00782-x;
RA Kim S., Kim J., Park S., Park J.J., Lee S.;
RT "Drosophila Graf regulates mushroom body beta-axon extension and olfactory
RT long-term memory.";
RL Mol. Brain 14:73-73(2021).
CC -!- FUNCTION: GTPase-activating protein for Rho family proteins
CC (PubMed:33835025). Essential component of the CLIC (clathrin-
CC independent carrier)/GEEC (GPI-anchored protein-enriched early
CC endocytic compartment) endocytic pathway (PubMed:28993397). During
CC hematopoiesis, inhibits Egfr-ras-MAPK signaling by promoting Spi-
CC induced Egfr internalization through CLIC/GEEC endocytosis, thereby
CC preventing plasmatocyte overproliferation (PubMed:28993397). Essential
CC for normal mushroom body (MB) development and consequently the
CC formation of olfactory long-term memories (PubMed:33892766). During MD
CC development, required to stop the MB beta-lobe from crossing the brain
CC midline, possibly acting via its role in the CLIC/GEEC endocytic
CC pathway to down-regulate the Egfr-ras-MAPK signaling at the tip of the
CC beta-lobes (PubMed:33892766). Required during embryo cellularization
CC for maintaining and regulating the rate of actomyosin ring constriction
CC (PubMed:33835025). During cellularization, inhibits Rho-GTP levels at
CC the furrow canal tip in a spatiotemporal manner, thus delaying the
CC onset of actomyosin contraction and ensuring appropriate closure of the
CC cells at the base of nuclei after membrane extension (PubMed:33835025).
CC {ECO:0000269|PubMed:28993397, ECO:0000269|PubMed:33835025,
CC ECO:0000269|PubMed:33892766}.
CC -!- SUBUNIT: Interacts with Egfr (when ubiquitinated).
CC {ECO:0000269|PubMed:28993397}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol {ECO:0000269|PubMed:33835025}.
CC Cytoplasm {ECO:0000269|PubMed:28993397}. Cytoplasmic vesicle
CC {ECO:0000269|PubMed:28993397}. Note=During embryonic cellularization,
CC detected at the furrow canal tips with enrichment at the edges
CC (PubMed:33835025). Recruited to the furrow tips at early
CC cellularization, expression levels are highest during mid-
CC cellularization but then decrease at late cellularization when it
CC becomes predominately cytosolic (PubMed:33835025). In embryos and
CC larvae, displays a punctate localization in the cytoplasm of
CC plasmatocytes (PubMed:28993397). In larvae, also displays a punctate
CC localization in hemolymph crystal cells (PubMed:28993397). During Egfr
CC internalization, localizes with Egfr and spi, to punctate or tubular
CC structures connected to the cell surface (PubMed:28993397).
CC {ECO:0000269|PubMed:28993397, ECO:0000269|PubMed:33835025}.
CC -!- ALTERNATIVE PRODUCTS:
CC Event=Alternative splicing; Named isoforms=2;
CC Name=G {ECO:0000312|FlyBase:FBgn0030685}; Synonyms=H
CC {ECO:0000312|FlyBase:FBgn0030685}, I {ECO:0000312|FlyBase:FBgn0030685};
CC IsoId=X2JDY8-1; Sequence=Displayed;
CC Name=A {ECO:0000312|FlyBase:FBgn0030685}; Synonyms=D
CC {ECO:0000312|FlyBase:FBgn0030685}, E {ECO:0000312|FlyBase:FBgn0030685},
CC J {ECO:0000312|FlyBase:FBgn0030685};
CC IsoId=X2JDY8-2; Sequence=VSP_061214;
CC -!- TISSUE SPECIFICITY: In the adult brain, expressed in the antennal lobe,
CC the subesophageal ganglion and the alpha/beta neurons of the mushroom
CC body. {ECO:0000269|PubMed:33892766}.
CC -!- DEVELOPMENTAL STAGE: In the larval primary lymph gland, expressed
CC throughout the medullary and cortical zones (at protein level). Also
CC detected in larval and embryo plasmatocytes (at protein level).
CC {ECO:0000269|PubMed:28993397}.
CC -!- DOMAIN: The Rho-GAP domain is necessary for inhibiting contractile ring
CC constriction during cellularization. {ECO:0000269|PubMed:33835025}.
CC -!- DISRUPTION PHENOTYPE: RNAi-mediated knockdown in embryos is lethal at
CC 24 hr, likely due to defective cellularization (PubMed:33835025).
CC During cellularization, the F-actin network at the base of the furrow
CC canal is unorganized, the furrow tips contain wavy edges and there is a
CC loss of contact between adjacent furrow tips (PubMed:33835025).
CC Actomyosin ring constriction is enhanced, and the nuclei have a
CC bottleneck appearance likely due to premature constriction at the
CC furrow canal tip during the mid cellularization stage
CC (PubMed:33835025). Another study reports that flies are viable and
CC fertile (PubMed:28993397). RNAi-mediated knockdown in the alpha/beta
CC neurons of the adult mushroom body (MB), results in abnormal crossing
CC of the MB beta lobe over the brain midline (PubMed:33892766).
CC {ECO:0000269|PubMed:28993397, ECO:0000269|PubMed:33835025,
CC ECO:0000269|PubMed:33892766}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAO39581.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; AE014298; AAF48506.3; -; Genomic_DNA.
DR EMBL; AE014298; AAS65342.1; -; Genomic_DNA.
DR EMBL; AE014298; AAS65343.2; -; Genomic_DNA.
DR EMBL; AE014298; AAS65344.1; -; Genomic_DNA.
DR EMBL; AE014298; AAS65345.1; -; Genomic_DNA.
DR EMBL; AE014298; AAS65346.2; -; Genomic_DNA.
DR EMBL; AE014298; AHN59766.1; -; Genomic_DNA.
DR EMBL; BT003577; AAO39581.1; ALT_FRAME; mRNA.
DR RefSeq; NP_001285296.1; NM_001298367.1. [X2JDY8-1]
DR RefSeq; NP_573070.2; NM_132842.3. [X2JDY8-2]
DR RefSeq; NP_996441.2; NM_206718.2. [X2JDY8-1]
DR RefSeq; NP_996442.1; NM_206719.2. [X2JDY8-2]
DR RefSeq; NP_996443.1; NM_206720.2. [X2JDY8-2]
DR RefSeq; NP_996444.2; NM_206721.2. [X2JDY8-1]
DR RefSeq; NP_996445.1; NM_206722.2. [X2JDY8-2]
DR AlphaFoldDB; X2JDY8; -.
DR SMR; X2JDY8; -.
DR IntAct; X2JDY8; 8.
DR STRING; 7227.FBpp0073979; -.
DR PRIDE; X2JDY8; -.
DR EnsemblMetazoa; FBtr0074194; FBpp0073979; FBgn0030685. [X2JDY8-2]
DR EnsemblMetazoa; FBtr0074197; FBpp0089212; FBgn0030685. [X2JDY8-2]
DR EnsemblMetazoa; FBtr0074198; FBpp0089213; FBgn0030685. [X2JDY8-2]
DR EnsemblMetazoa; FBtr0340305; FBpp0309266; FBgn0030685. [X2JDY8-1]
DR EnsemblMetazoa; FBtr0340306; FBpp0309267; FBgn0030685. [X2JDY8-1]
DR EnsemblMetazoa; FBtr0340307; FBpp0309268; FBgn0030685. [X2JDY8-1]
DR EnsemblMetazoa; FBtr0345133; FBpp0311354; FBgn0030685. [X2JDY8-2]
DR GeneID; 32522; -.
DR KEGG; dme:Dmel_CG8948; -.
DR UCSC; CG8948-RA; d. melanogaster.
DR CTD; 32522; -.
DR FlyBase; FBgn0030685; Graf.
DR VEuPathDB; VectorBase:FBgn0030685; -.
DR eggNOG; KOG1451; Eukaryota.
DR GeneTree; ENSGT00940000168384; -.
DR HOGENOM; CLU_011532_2_0_1; -.
DR OMA; LAXIFNT; -.
DR OrthoDB; 693048at2759; -.
DR Reactome; R-DME-8980692; RHOA GTPase cycle.
DR Reactome; R-DME-9013026; RHOB GTPase cycle.
DR Reactome; R-DME-9013106; RHOC GTPase cycle.
DR Reactome; R-DME-9013148; CDC42 GTPase cycle.
DR Reactome; R-DME-9013149; RAC1 GTPase cycle.
DR Reactome; R-DME-9013405; RHOD GTPase cycle.
DR BioGRID-ORCS; 32522; 0 hits in 3 CRISPR screens.
DR ChiTaRS; Graf; fly.
DR GenomeRNAi; 32522; -.
DR Proteomes; UP000000803; Chromosome X.
DR Bgee; FBgn0030685; Expressed in spermathecum and 26 other tissues.
DR ExpressionAtlas; X2JDY8; baseline and differential.
DR GO; GO:1903144; C:actomyosin contractile ring actin filament; IDA:UniProtKB.
DR GO; GO:0110071; C:cellularization cleavage furrow invagination front; IDA:UniProtKB.
DR GO; GO:0031410; C:cytoplasmic vesicle; IDA:FlyBase.
DR GO; GO:0005829; C:cytosol; IDA:UniProtKB.
DR GO; GO:0005096; F:GTPase activator activity; IMP:UniProtKB.
DR GO; GO:0005543; F:phospholipid binding; ISS:FlyBase.
DR GO; GO:0140036; F:ubiquitin-dependent protein binding; IDA:FlyBase.
DR GO; GO:0036089; P:cleavage furrow formation; IMP:UniProtKB.
DR GO; GO:0042059; P:negative regulation of epidermal growth factor receptor signaling pathway; IMP:FlyBase.
DR GO; GO:0002092; P:positive regulation of receptor internalization; IMP:FlyBase.
DR GO; GO:0031991; P:regulation of actomyosin contractile ring contraction; IMP:UniProtKB.
DR GO; GO:0007266; P:Rho protein signal transduction; ISM:FlyBase.
DR GO; GO:0110069; P:syncytial embryo cellularization; IMP:UniProtKB.
DR Gene3D; 1.10.555.10; -; 1.
DR Gene3D; 1.20.1270.60; -; 1.
DR Gene3D; 2.30.29.30; -; 1.
DR InterPro; IPR027267; AH/BAR_dom_sf.
DR InterPro; IPR011993; PH-like_dom_sf.
DR InterPro; IPR001849; PH_domain.
DR InterPro; IPR008936; Rho_GTPase_activation_prot.
DR InterPro; IPR000198; RhoGAP_dom.
DR InterPro; IPR036028; SH3-like_dom_sf.
DR InterPro; IPR001452; SH3_domain.
DR Pfam; PF00169; PH; 1.
DR Pfam; PF00620; RhoGAP; 1.
DR Pfam; PF14604; SH3_9; 1.
DR SMART; SM00324; RhoGAP; 1.
DR SMART; SM00326; SH3; 1.
DR SUPFAM; SSF103657; SSF103657; 1.
DR SUPFAM; SSF48350; SSF48350; 1.
DR SUPFAM; SSF50044; SSF50044; 1.
DR PROSITE; PS50003; PH_DOMAIN; 1.
DR PROSITE; PS50238; RHOGAP; 1.
DR PROSITE; PS50002; SH3; 1.
PE 1: Evidence at protein level;
KW Alternative splicing; Coiled coil; Cytoplasm; Cytoplasmic vesicle;
KW GTPase activation; Reference proteome; SH3 domain.
FT CHAIN 1..1025
FT /note="Rho GTPase-activating protein Graf"
FT /id="PRO_0000453923"
FT DOMAIN 271..388
FT /note="PH"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00145"
FT DOMAIN 402..589
FT /note="Rho-GAP"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00172"
FT DOMAIN 963..1023
FT /note="SH3"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00192"
FT REGION 824..866
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 847..866
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT VAR_SEQ 284..293
FT /note="SSILKISLLE -> K (in isoform A)"
FT /id="VSP_061214"
FT MUTAGEN 28..1025
FT /note="Missing: In Graf-CR57; semi-embryonic lethal, with
FT 71% of embryos dying at 24 hr. Displays cellularization
FT defects including the loss of polygonal F-actin
FT architecture and premature actomyosin ring formation during
FT early cellularization, and enhanced ring constriction
FT during mid and late cellularization. Nuclei also have a
FT bottleneck appearance likely due to the premature ring
FT constriction at the furrow canal tip the mid
FT cellularization stage."
FT /evidence="ECO:0000269|PubMed:33835025"
FT MUTAGEN 393..588
FT /note="Missing: During cellularization, unable to rescue
FT increased ring constriction in Graf-CR57 mutants."
FT /evidence="ECO:0000269|PubMed:33835025"
SQ SEQUENCE 1025 AA; 115670 MW; 911DE93A56E4F4FE CRC64;
MGGGKNVRRG LEPLEFEECI VDSPDFRENL NRHEKELDHT SHQIKRIIKE VKDLMSAAKI
LSTRMKQLAI LLNDFNFECI GTAQTDDENV ICESLKRFGA IIGNIEDERE KMLTLADKHI
IESLEDFRKK QIGGVKENKK KFDKKTEKFC QSQERFLNMS TKKPENTIQE ADASLGMHER
EYIQESLSYV LRIQEVQERI KFEFVEILLA FISGWLVFYH TAHEQAEDHR DYLQDLRHKV
QKTRENFEEA REKVTELKTK YMEKRTKPEE IFTKRGYLFL MEKSSILKIS LLEPFKATWT
KYYCTFKKQK REFTMLQFNQ MNHNFTRPEA RDDEKLTLFS CQRRASEFEK RFCFDLTFKE
KPGVVYTFQA LSEKDHRYWI SAMDGTEPTY LAPGKIKVSE AYHLDEAGFM FIRRCIQVLE
IRGLEDEGIY RKSGVGTKIS KLLALGLNQK ESDDVFVDDK YRDLMESNTI ASALKMYLRN
LNEPLMTYQY HSDFIEAAKQ ETLNQRVNEV HKLVYKLPQP NFQMLDMVIC HLTDVSRKYE
KNKMSVFNLG VVFGPTLLRP REESVAAILD IKFNNIVINI LIDNYERIFK NKPSADIKLP
DATKAPSIMY PSRSSPPTRM PRASQIGKAA STGAMGSSSN TAGNPMFLNQ QKIYRVVSKT
NCTEPTMSSS LQNIPNGDNY ALGSNAMNSS GGAQCISLSP PMHMLNGILS PTIGSINNLH
TISKNETSTR RYVPDTEIAP DYGIIGANNG GVTLQSHHAV PVSSTSNFRH PEYLMTTANP
VTQSGSSSHI YTNTSSAGAN SSNRISLSNV SPPNTAMRKE RFLGSASGPQ QHPPVQRGLH
SYGQTKHYSP LMPTSTSSSN DSVCDSLSSN NGLGSSIVAN SGNSGNVAQH LSARNTNDYA
LISSQNSSLS PHLGPTCDEV VTATTLPSVS LSCGGTASES TEYPPSKMHR NRDVNQIKRD
LSTGTARVRT LYACMGESEG ELSFEPNQII TNVRYSHEPG WLQGTLNGKT GLIPENYVEH
LKPHH
