RhoA prenylation is required for promotion of cell growth and transformation and cytoskeleton organization but not for induction of serum response element transcription.
The importance of post-translational geranylgeranylation of the GTPase RhoA for its ability to induce cellular proliferation and malignant transformation is not well understood. In this manuscript we demonstrate that geranylgeranylation is required for the proper cellular localization of V14RhoA and for its ability to induce actin stress fiber and focal adhesion formation. Furthermore, V14RhoA geranylgeranylation was also required for suppressing p21(WAF) transcription, promoting cell cycle progression and cellular proliferation. The ability of V14RhoA to induce focus formation and enhance plating efficiency and oncogenic Ras anchorage-dependent growth was also dependent on its geranylgeranylation. The only biological activity of V14RhoA that was not dependent on its prenylation was its ability to induce serum response element transcriptional activity. Furthermore, we demonstrate that a farnesylated form of V14RhoA was also able to bind RhoGDI-1, was able to induce cytoskeleton organization, proliferation, and transformation, and was just as potent as geranylgeranylated V14RhoA at suppressing p21(WAF) transcriptional activity. These results demonstrate that RhoA geranylgeranylation is required for its biological activity and that the nature of the lipid modification is not critical.
Allal
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Oncologie Cellulaire et Moléculaire, EA 2048 Université Paul Sabatier, Centre de Lutte Contre le Cancer Claudius Regaud, 20-24 rue du Pont Saint-Pierre, 31052 Toulouse cedex, France.
Favre
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Couderc
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Salicio
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Sixou
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Hamilton
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AD
Sebti
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Lajoie-Mazenc
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Pradines
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D016475:3T3 Cells; D000199:Actins / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002453:Cell Cycle; D002455:Cell Division; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism* / Q000502:physiology; D004268:DNA-Binding Proteins; D003902:Detergents / Q000494:pharmacology; D022001:Focal Adhesions / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D011905:Genes, ras / Q000235:genetics; D005982:Glutathione Transferase / Q000378:metabolism; D050356:Lipid Metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009687:Nuclear Proteins; D017830:Octoxynol; D010957:Plasmids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D017368:Protein Prenylation; D011499:Protein Processing, Post-Translational; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020218:Response Elements / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D026362:Serum Response Factor; D022502:Stress Fibers / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D016596:Vinculin / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D000199:Actins; C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004268:DNA-Binding Proteins; D003902:Detergents; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D026362:Serum Response Factor; D016596:Vinculin; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D005982:Glutathione Transferase; D020558:GTP Phosphohydrolases; D016283:Proto-Oncogene Proteins p21(ras); D020742:rhoA GTP-Binding Protein