JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N3_FDR10.csvhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N1_FDR10.csvhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N2_FDR10.csvhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N2.rawhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N1.rawhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N3.rawhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N2_FDR10.mzidhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N1_FDR10.mzidhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N3_FDR10.mzidhttps://storage.jpostdb.org/JPST002477/files/Elmo1_Dock5_Rac1_RhoG_20210318.fastahttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N2.mgfhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N1.mgfhttps://storage.jpostdb.org/JPST002477/files/SN210154_E_7ul_N3.mgfprimaryOK200ProteomicsMikako ShirouzuHomo Sapiens (human)https://repository.jpostdb.org/entry/JPST002477RIKEN BDRjPOSTThe dedicator of cytokinesis (DOCK)/engulfment and cell motility (ELMO) complex serves as a guanine nucleotide exchange factor (GEF) for the GTPase Rac. RhoG, another GTPase, activates the ELMO-DOCK-Rac pathway during engulfment and migration. Recent cryo-EM structures of the DOCK2/ELMO1 and DOCK2/ELMO1/Rac1 complexes have identified closed and open conformations that are key to understanding the autoinhibition mechanism. Nevertheless, the structural details of RhoG-mediated activation of the DOCK/ELMO complex remain elusive. Herein, we present cryo-EM structures of DOCK5/ELMO1 alone and in complex with RhoG and Rac1. The DOCK5/ELMO1 structure exhibits a closed conformation similar to that of DOCK2/ELMO1, suggesting a shared regulatory mechanism of the autoinhibitory state across DOCK-A/B subfamilies (DOCK1-5). Conversely, the RhoG/DOCK5/ELMO1/Rac1 complex adopts an open conformation that differs from that of the DOCK2/ELMO1/Rac1 complex, with RhoG binding to both ELMO1 and DOCK5. The alignment of the DOCK5 phosphatidylinositol (3,4,5)-trisphosphate binding site with the RhoG C-terminal lipidation site suggests simultaneous binding of RhoG and DOCK5/ELMO1 to the plasma membrane. Structural comparison of the apo and RhoG-bound states revealed that RhoG facilitates a closed-to-open state conformational change of DOCK5/ELMO1. Biochemical and surface plasmon resonance (SPR) assays confirm that RhoG enhances the Rac GEF activity of DOCK5/ELMO1 and increases its binding affinity for Rac1. Further analysis of structural variability underscored the conformational flexibility of the DOCK5/ELMO1/Rac1 complex core, potentially facilitating the proximity of the DOCK5 GEF domain to the plasma membrane. These findings elucidate the structural mechanism underlying the RhoG-induced allosteric activation and membrane binding of the DOCK/ELMO complex.RhoG facilitates a conformational transition in the guanine nucleotide exchange factor complex DOCK5/ELMO1 to an open state.Kukimoto-Niino Mutsuko M, Katsura Kazushige K, Ishizuka-Katsura Yoshiko Y, Mishima-Tsumagari Chiemi C, Yonemochi Mayumi M, Inoue Mio M, Nakagawa Reiko R, Kaushik Rahul R, Zhang Kam Y J KYJ, Shirouzu Mikako MRIB1, Rac GTPase, DracA, dRac1, E130320D18, AL023026, 6330578D22Rik, D-Rac1, DRac, 1110060D06Rik, Drac1, elmo-1, CED12, AI956923, rac, RAC1, D-Rac 1, ced-12, p21-Rac1, GTPase Rac1, rac1, Rac-1, D5Ertd559e, D-Rac, ELMO1s, dRac, ced12, AI666732, C230095H21Rik, Dm Rac1, DmelCG2248, DmRAC1, KIAA0281, MIG5, 2810426G09Rik, 2248, TC25, Arhg., DmRAC, RNS1, Drac1a, arhg, TC-25, BC016533, DRac1, rlc, RacA, CED-12, lr2, ARHG, Drac, Rac, DRacA, Dmrac1, Sid10750, ELMO-1, CG2248ced12, GDP Dissociation Factor, Guanine Nucleotide Releasing Factors, AI666732, GDP Dissociation Stimulators, C230095H21Rik, Factors, GDP Exchange, E130320D18, 6330578D22Rik, KIAA0281., 2810426G09Rik, Arhg, GDP-GTP Reversing, GDP, 1110060D06Rik, Exchange Factors, GDP-GTP Reversing Factors, AI956923, elmo-1, CED12, Guanine Nucleotide Releasing Factor, arhg, GDP-GTP Exchange Protein, ced-12, BC016533, reaction, rlc, GDP-GTP, GDP GTP Reversing Factors, CED-12, lr2, ARHG, Reversing Factors, Guanine Nucleotide Exchange Factor, GDP GTP Exchange Protein, guanine nucleotide exchange factor, Guanine-Nucleotide-Releasing Factor, Sid10750, ELMO1s, ELMO-1, GDP Exchange FactorsRac GTPase, PRKBA, Plasma Membranes, xspr, Akt/PKB, Huntington disease gene regulatory region-binding protein 1, Lfc, Activity, determination, D-Rac1, D-Rac2, DAKT1/PKB, DOCK, CED12, rac, l(2)04723, GEFH1, AKT1, PtdIns, Ced12, Ras guanyl-nucleotide exchange factor activity, Divisions, ced5, DOCK180, Cytokineses, AA408978, Membrane Tissues, Readability, GTPases, DmelCG8556, Rac-2, Rac-1, 9130006G06Rik, Reversing Factors, symptoms, cell movement, Cell Motility, IKKg, KEY, Key, ced12, GDP Dissociation Stimulators, C230095H21Rik, Sar guanyl-nucleotide exchange factor activity, GDP Exchange, AI327394, KIAA0281, Surface, membrane region, Triphosphate Phosphohydrolases, Tissue, Cell Locomotion, CG3727, PKB|Akt, GDP-GTP Exchange Protein, RacPK, Phosphatidyl Inositol, SDR38C1, inner endospore membrane, Inositol Phospholipids, Movements, plasma membrane lipid bilayer, cytokinesis involved in cell cycle, Plasmon Resonances, plasmalemma, dced-12, GDP GTP Reversing Factors, Plasma Membrane, Locomotion, Dock, guanine nucleotide exchange factor, 1.1.1.153, RIB1, screening, Membrane Tissue, Motility, Guanosine, dRac1, Elmo, dakt, E130320D18, TAC1R, GDP-GTP Reversing, GTP Phosphohydrolase, Cell Migration, 1110060D06Rik, Exchange Factors, integral to membrane, CG8556, DelGEF, AI227039, l(3)89Bq, CED-5, NK1R, Tac1r, PBL, Pbl, RAC1, D630004B07Rik, gtl11, D-Rac 1, D-Rac 2, Binding, p21-Rac1, Rho guanyl-nucleotide exchange factor activity, PKB/Akt, PKB/AKT, AKT/PKB, DmIKKgamma, cellular membrane, elmo, GTPase Rac1, dAkt, dAKT, rac1, ELMO, rac2, dIKK, Hch, GDP GTP Exchange Protein, Kenny, p-Akt, bacterial inner membrane, Phosphoglyceride, DAkt, GEF-H1, ELMO1s, activation, region, l(3)04226, Si-1-2, AI662014, DPKB, AI666732, Si-1-2-19, Dm Rac1, cell cycle cytokinesis, Factors, cell membrane, DmRAC2, pAkt, DmRAC1, phosphatidylinositols, P40, 2810426G09Rik, IKK-gamma, 2248, signs, TC25, Ran guanyl-nucleotide exchange factor activity, dAKT/dPKB, PKB/dAKT, Nk1r, arhg, 0542/03, TC-25, GLUT4 enhancer factor, Phosphoinositides, Phosphohydrolase, DRac1, Dakt, DRac2, DmelCG16910, RacB, RacA, dPKB, ARHG, Inositide Phospholipids, DmelCG3727, ENSMUSG00000071345, Guanine-Nucleotide-Releasing Factor, DRacA, DRacB, mKIAA1478, Plasmon Resonance, Dmrac1, Resonance, guanyl-nucleotide releasing factor, RAC-ALPHA, GTP, DRAC-PK85, DracA, Dced-12, DmelCG4006, Inositide, Migration, DmelCG5336, AL023026, conformation, Arhg, DRac, Cytoplasmic Membrane, Drac2, Phosphatidylinositol, GDP, Drac1, GDP-GTP Reversing Factors, CG8114, GDS, AW122239, dIKK-gamma, ARF guanyl-nucleotide exchange factor activity, Guanosine Triphosphate, ced-12, Dpkb, GDP-GTP, GEF, Gef, Surface Plasmon Resonances, integral component of membrane, DmIKK-gamma, cell locomotion, Sites, HDBP1, Phosphatidyl, dmIKKgamma, Nck, D-Rac, IKK[[gamma]], Inositide Phospholipid, doc, cAMP-dependent guanyl-nucleotide exchange factor activity, dRac, Site, Guanosine Triphosphate Phosphohydrolases, Guanine Nucleotide Releasing Factors, GNRP, Phospholipid, anon-WO0172774.22, Tissues, Flexibility, MIG5, ligand, MBC, anon-WO0172774.25, Gm10328, Division, DmRAC, ECT2, RNS1, LFP40, Cell Movements, Cytoplasmic, Ral guanyl-nucleotide exchange factor activity, BC016533, Movement, Combining Site, AA409688, IKK, rlc, AKT, Akt, Cytoplasmic Membranes, guanyl-nucleotide release factor activity, lr2, Guanine Nucleotide Exchange Factor, DRAC-PK, Drac, region of membrane, Band25, site, Sid10750, Combining Sites, relational structural quality, CG2248, integral to plasma membrane, GTPase, movement of a cell, GDP Dissociation Factor, NKIR, membrane, Inositol Phosphoglyceride, findings, Guanosinetriphosphatases, 6330578D22Rik, akt, AI854900, integral component of plasma membrane, Dm0447, 0083/20, elmo-1, AI956923, SPR, Spr, Gnefr, DAkt1, DAKT1, Phosphoglycerides, Rab guanyl-nucleotide exchange factor activity, Binding Site, Cell, l(3)09645, Inositol, IKKgamma, DmelCG8114, PKB, CWS6, Inositol Phosphoglycerides, D5Ertd559e, Cytoplasmic Division, chemical analysis, PKB-ALPHA, whole membrane, core, D-Akt, CG5336, CG4006, Rap guanyl-nucleotide exchange factor activity, Cell Membranes, GDP-dissociation stimulator activity, Dakt1, Plasma, MgcRacGAP, Phosphoinositide, Membranes, Resonances, transmembrane, Gnef, DmelCG2248, akt1, Phospholipids, dAkt1, Surface Plasmon, Dmikkgamma, dck, Lbcl1, DRAC-PK66, dced-12., HDBP-1, Combining, Guanine Nucleotide Releasing Factor, Understanding, Membrane, mKIAA0651, CG16910, Drac1b, Drac1a, Phosphohydrolases, 0293/09, dAkt/PKB, Rac guanyl-nucleotide exchange factor activity, dakt1, Inositol Phospholipid, CED-12, PKBalpha, Cytoplasmic Divisions, dAKT1, Rac1b, RAC, Rac, assay, cytoplasmic membrane, PI, General activity, ELMO-1, GDP Exchange FactorsRIB1, Rac GTPase, DracA, dRac1, E130320D18, AL023026, 6330578D22Rik, D-Rac1, Arhg, DRac, 1110060D06Rik, Drac1, elmo-1, CED12, AI956923, rac, Spectrum Analyses, RAC1, Spectrum Analysis, D-Rac 1, Spectroscopy, ced-12, p21-Rac1, MS, GTPase Rac1, rac1, Rac-1, D5Ertd559e, Mass, Analysis, D-Rac, ELMO1s, Mass Spectrum Analyses, Mass Spectroscopy, dRac, Mass Spectrum Analysis, ced12, Mass Spectrum, AI666732, C230095H21Rik, Dm Rac1, DmelCG2248, DmRAC1, Analyses, KIAA0281, MIG5, 2810426G09Rik, Spectrometry, 2248, TC25, DmRAC, RNS1, Drac1a, arhg, TC-25, BC016533, DRac1, rlc, RacA, CED-12, lr2, ARHG, Drac, BS3., Rac, DRacA, Dmrac1, Sid10750, ELMO-1, CG2248falseCrosslinking MS of Elmo1,Dock5,Rac1 and RhoG complexesCrosslink mass spectrometry of Elmo1,Dock5,Rac1 and RhoG complexes using BS3 crosslinker.Sat Jan 18 00:00:00 GMT 2025PXD048862960638857861