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| | ==Farnesylated RhoA-GDP in complex with RhoGDI-alpha, lysine acetylated at K178== | | ==Farnesylated RhoA-GDP in complex with RhoGDI-alpha, lysine acetylated at K178== |
| - | <StructureSection load='5fr2' size='340' side='right' caption='[[5fr2]], [[Resolution|resolution]] 3.35Å' scene=''> | + | <StructureSection load='5fr2' size='340' side='right'caption='[[5fr2]], [[Resolution|resolution]] 3.35Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5fr2]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5FR2 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5FR2 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5fr2]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Bos_taurus Bos taurus] and [https://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5FR2 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5FR2 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=BGC:BETA-D-GLUCOSE'>BGC</scene>, <scene name='pdbligand=FAR:FARNESYL'>FAR</scene>, <scene name='pdbligand=GDP:GUANOSINE-5-DIPHOSPHATE'>GDP</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 3.35Å</td></tr> |
| - | <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=ALY:N(6)-ACETYLLYSINE'>ALY</scene></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ALY:N(6)-ACETYLLYSINE'>ALY</scene>, <scene name='pdbligand=BGC:BETA-D-GLUCOSE'>BGC</scene>, <scene name='pdbligand=FAR:FARNESYL'>FAR</scene>, <scene name='pdbligand=GDP:GUANOSINE-5-DIPHOSPHATE'>GDP</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5fr1|5fr1]]</td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5fr2 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5fr2 OCA], [https://pdbe.org/5fr2 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5fr2 RCSB], [https://www.ebi.ac.uk/pdbsum/5fr2 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5fr2 ProSAT]</span></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5fr2 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5fr2 OCA], [http://pdbe.org/5fr2 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5fr2 RCSB], [http://www.ebi.ac.uk/pdbsum/5fr2 PDBsum]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/RHOA_HUMAN RHOA_HUMAN]] Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion. Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague, and Yersinia pseudotuberculosis, which causes gastrointestinal disorders. Stimulates PKN2 kinase activity. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization.<ref>PMID:8910519</ref> <ref>PMID:9121475</ref> <ref>PMID:12900402</ref> <ref>PMID:16103226</ref> <ref>PMID:16236794</ref> <ref>PMID:19934221</ref> <ref>PMID:20937854</ref> <ref>PMID:20974804</ref> [[http://www.uniprot.org/uniprot/GDIR1_BOVIN GDIR1_BOVIN]] In glioma cells, inhibits cell migration and invasion by mediating the signals of SEMA5A and PLXNB3 that lead to inactivation of RAC1 (By similarity). Regulates the GDP/GTP exchange reaction of the Rho proteins by inhibiting the dissociation of GDP from them, and the subsequent binding of GTP to them.<ref>PMID:2120668</ref> <ref>PMID:9194563</ref> | + | [https://www.uniprot.org/uniprot/RHOA_HUMAN RHOA_HUMAN] Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion. Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague, and Yersinia pseudotuberculosis, which causes gastrointestinal disorders. Stimulates PKN2 kinase activity. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization.<ref>PMID:8910519</ref> <ref>PMID:9121475</ref> <ref>PMID:12900402</ref> <ref>PMID:16103226</ref> <ref>PMID:16236794</ref> <ref>PMID:19934221</ref> <ref>PMID:20937854</ref> <ref>PMID:20974804</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
| - | Rho proteins are major regulators of the cytoskeleton. As most Ras-related proteins, they switch between an active, GTP-bound and an inactive, GDP-bound conformation. Rho proteins are targeted to the plasma membrane via a polybasic region and a prenyl group attached to a C-terminal cysteine residue. To distribute Rho proteins in the cell, the molecular chaperone RhoGDIalpha binds to the prenylated Rho proteins forming a cytosolic pool of mainly GDP-loaded Rho. Most studies characterized the interaction of prenylated Rho proteins and RhoGDIalpha. However, RhoGDIalpha was also shown to bind to nonprenylated Rho proteins with physiologically relevant micomolar affinities. Recently, it was discovered that RhoGDIalpha is targeted by post-translational lysine acetylation. For one site, K141, it was hypothesized that acetylation might lead to increased levels of formation of filamentous actin and filopodia in mammalian cells. The functional consequences of lysine acetylation for the interplay with nonprenylated RhoA have not been investigated. Here, we report that lysine acetylation at lysines K127 and K141 in the RhoGDIalpha immunoglobulin domain interferes with the interaction toward nonprenylated RhoA using a combined biochemical and biophysical approach. We determined the first crystal structure of a doubly acetylated protein, RhoGDIalpha, in complex with RhoA.GDP. We discover that the C-terminus of RhoA adopts a different conformation forming an intermolecular beta-sheet with the RhoGDIalpha immunoglobulin domain. | + | Rho-proteins are small GTP/GDP-binding proteins primarily involved in cytoskeleton regulation. Their GTP/GDP-cycle is often tightly connected to a membrane/cytosol-cycle regulated by the Rho guanine nucleotide dissociation inhibitor alpha (RhoGDIalpha). RhoGDIalpha has been regarded as a housekeeping regulator essential to control homeostasis of Rho-proteins. Recent proteomic screens showed that RhoGDIalpha is extensively lysine acetylated. Here, we present the first comprehensive structural and mechanistic study to show how RhoGDIalpha function is regulated by lysine acetylation. We discover that lysine acetylation impairs Rho-protein binding and increases GEF-catalysed nucleotide exchange on RhoA, being prerequisites to constitute a bona fide GDI-displacement factor. RhoGDIalpha-acetylation interferes with Rho-signalling resulting in alteration of cellular filamentous actin. Finally, we discover that RhoGDIalpha is endogenously acetylated in mammalian cells, we identify CBP, p300, pCAF as RhoGDIalpha-acetyltransferases and Sirt2, HDAC6 as specific deacetylases, showing the biological significance of this post-translational modification. |
| | | | |
| - | RhoGDIalpha Acetylation at K127 and K141 Affects Binding toward Nonprenylated RhoA.,Kuhlmann N, Wroblowski S, Scislowski L, Lammers M Biochemistry. 2016 Jan 4. PMID:26695096<ref>PMID:26695096</ref>
| + | Structural and mechanistic insights into the regulation of the fundamental Rho-regulator RhoGDIalpha by lysine acetylation.,Kuhlmann N, Wroblowski S, Knyphausen P, de Boor S, Brenig J, Zienert AY, Meyer-Teschendorf K, Praefcke GJ, Nolte H, Kruger M, Schacherl M, Baumann U, James LC, Chin JW, Lammers M J Biol Chem. 2015 Dec 30. pii: jbc.M115.707091. PMID:26719334<ref>PMID:26719334</ref> |
| | | | |
| | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| | <div class="pdbe-citations 5fr2" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 5fr2" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Guanine nucleotide dissociation inhibitor|Guanine nucleotide dissociation inhibitor]] |
| | + | *[[Rho GTPase 3D structures|Rho GTPase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Baumann, U]] | + | [[Category: Bos taurus]] |
| - | [[Category: Boor, S de]] | + | [[Category: Homo sapiens]] |
| - | [[Category: Brenig, J]] | + | [[Category: Large Structures]] |
| - | [[Category: Chin, J W]] | + | [[Category: Baumann U]] |
| - | [[Category: James, L C]] | + | [[Category: Brenig J]] |
| - | [[Category: Knyphausen, P]] | + | [[Category: Chin JW]] |
| - | [[Category: Krueger, M]] | + | [[Category: James LC]] |
| - | [[Category: Kuhlmann, N]] | + | [[Category: Knyphausen P]] |
| - | [[Category: Lammers, M]] | + | [[Category: Krueger M]] |
| - | [[Category: Meyer-Teschendorf, K]] | + | [[Category: Kuhlmann N]] |
| - | [[Category: Nolte, H]] | + | [[Category: Lammers M]] |
| - | [[Category: Praefcke, G J.K]] | + | [[Category: Meyer-Teschendorf K]] |
| - | [[Category: Schacherl, M]] | + | [[Category: Nolte H]] |
| - | [[Category: Wroblowski, S]] | + | [[Category: Praefcke GJK]] |
| - | [[Category: Zienert, A Y]] | + | [[Category: Schacherl M]] |
| - | [[Category: Cytoskeleton]] | + | [[Category: Wroblowski S]] |
| - | [[Category: Gdp]]
| + | [[Category: Zienert AY]] |
| - | [[Category: Lysine-acetylation]]
| + | [[Category: De Boor S]] |
| - | [[Category: Ras-superfamily]]
| + | |
| - | [[Category: Rhoa]]
| + | |
| - | [[Category: Rhogdi]]
| + | |
| - | [[Category: Signaling protein]]
| + | |
| Structural highlights
5fr2 is a 2 chain structure with sequence from Bos taurus and Homo sapiens. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
| | Method: | X-ray diffraction, Resolution 3.35Å |
| Ligands: | , , , , |
| Resources: | FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT |
Function
RHOA_HUMAN Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion. Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague, and Yersinia pseudotuberculosis, which causes gastrointestinal disorders. Stimulates PKN2 kinase activity. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization.[1] [2] [3] [4] [5] [6] [7] [8]
Publication Abstract from PubMed
Rho-proteins are small GTP/GDP-binding proteins primarily involved in cytoskeleton regulation. Their GTP/GDP-cycle is often tightly connected to a membrane/cytosol-cycle regulated by the Rho guanine nucleotide dissociation inhibitor alpha (RhoGDIalpha). RhoGDIalpha has been regarded as a housekeeping regulator essential to control homeostasis of Rho-proteins. Recent proteomic screens showed that RhoGDIalpha is extensively lysine acetylated. Here, we present the first comprehensive structural and mechanistic study to show how RhoGDIalpha function is regulated by lysine acetylation. We discover that lysine acetylation impairs Rho-protein binding and increases GEF-catalysed nucleotide exchange on RhoA, being prerequisites to constitute a bona fide GDI-displacement factor. RhoGDIalpha-acetylation interferes with Rho-signalling resulting in alteration of cellular filamentous actin. Finally, we discover that RhoGDIalpha is endogenously acetylated in mammalian cells, we identify CBP, p300, pCAF as RhoGDIalpha-acetyltransferases and Sirt2, HDAC6 as specific deacetylases, showing the biological significance of this post-translational modification.
Structural and mechanistic insights into the regulation of the fundamental Rho-regulator RhoGDIalpha by lysine acetylation.,Kuhlmann N, Wroblowski S, Knyphausen P, de Boor S, Brenig J, Zienert AY, Meyer-Teschendorf K, Praefcke GJ, Nolte H, Kruger M, Schacherl M, Baumann U, James LC, Chin JW, Lammers M J Biol Chem. 2015 Dec 30. pii: jbc.M115.707091. PMID:26719334[9]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Quilliam LA, Lambert QT, Mickelson-Young LA, Westwick JK, Sparks AB, Kay BK, Jenkins NA, Gilbert DJ, Copeland NG, Der CJ. Isolation of a NCK-associated kinase, PRK2, an SH3-binding protein and potential effector of Rho protein signaling. J Biol Chem. 1996 Nov 15;271(46):28772-6. PMID:8910519
- ↑ Vincent S, Settleman J. The PRK2 kinase is a potential effector target of both Rho and Rac GTPases and regulates actin cytoskeletal organization. Mol Cell Biol. 1997 Apr;17(4):2247-56. PMID:9121475
- ↑ Wing MR, Snyder JT, Sondek J, Harden TK. Direct activation of phospholipase C-epsilon by Rho. J Biol Chem. 2003 Oct 17;278(42):41253-8. Epub 2003 Aug 4. PMID:12900402 doi:http://dx.doi.org/10.1074/jbc.M306904200
- ↑ Yuce O, Piekny A, Glotzer M. An ECT2-centralspindlin complex regulates the localization and function of RhoA. J Cell Biol. 2005 Aug 15;170(4):571-82. PMID:16103226 doi:10.1083/jcb.200501097
- ↑ Kamijo K, Ohara N, Abe M, Uchimura T, Hosoya H, Lee JS, Miki T. Dissecting the role of Rho-mediated signaling in contractile ring formation. Mol Biol Cell. 2006 Jan;17(1):43-55. Epub 2005 Oct 19. PMID:16236794 doi:10.1091/mbc.E05-06-0569
- ↑ Bristow JM, Sellers MH, Majumdar D, Anderson B, Hu L, Webb DJ. The Rho-family GEF Asef2 activates Rac to modulate adhesion and actin dynamics and thereby regulate cell migration. J Cell Sci. 2009 Dec 15;122(Pt 24):4535-46. doi: 10.1242/jcs.053728. Epub 2009, Nov 24. PMID:19934221 doi:10.1242/jcs.053728
- ↑ Zaoui K, Benseddik K, Daou P, Salaun D, Badache A. ErbB2 receptor controls microtubule capture by recruiting ACF7 to the plasma membrane of migrating cells. Proc Natl Acad Sci U S A. 2010 Oct 26;107(43):18517-22. doi:, 10.1073/pnas.1000975107. Epub 2010 Oct 11. PMID:20937854 doi:10.1073/pnas.1000975107
- ↑ Wallace SW, Magalhaes A, Hall A. The Rho target PRK2 regulates apical junction formation in human bronchial epithelial cells. Mol Cell Biol. 2011 Jan;31(1):81-91. doi: 10.1128/MCB.01001-10. Epub 2010 Oct 25. PMID:20974804 doi:10.1128/MCB.01001-10
- ↑ Kuhlmann N, Wroblowski S, Knyphausen P, de Boor S, Brenig J, Zienert AY, Meyer-Teschendorf K, Praefcke GJ, Nolte H, Kruger M, Schacherl M, Baumann U, James LC, Chin JW, Lammers M. Structural and mechanistic insights into the regulation of the fundamental Rho-regulator RhoGDIalpha by lysine acetylation. J Biol Chem. 2015 Dec 30. pii: jbc.M115.707091. PMID:26719334 doi:http://dx.doi.org/10.1074/jbc.M115.707091
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