Interactions between Rho GTPases and Rho GDP dissociation inhibitor (Rho-GDI).
The Rho-GDP dissociation inhibitor (Rho-GDI) was used as bait in a two-hybrid screen of a human leucocyte cDNA library. Most of the isolated cDNAs encoded GTPases of the Rho subfamily: RhoA, B, C, Rac1, 2, CDC42 and RhoG. The newly discovered RhoH interacted very poorly with Rho-GDI. Another protein partner ... shared a homology with RhoA that points to Asp67(RhoA)-Arg68(RhoA)-Leu69(RhoA) as critical for interaction with Rho-GDI. A second screen with RhoA as bait led to the isolation of GDI only. In order to investigate the relative role of protein-protein and protein-lipid interactions between Rho GTPases and Rho-GDI, CAAX box mutants of RhoA were produced. They were found to interact with Rho-GDI as efficiently as wild type RhoA, indicating that protein-protein interactions alone lead to strong binding of the two proteins. The C-terminal polybasic region of RhoA was also shown to be a site of protein-protein interaction with Rho-GDI.
Mesh Terms:
Amino Acid Sequence, DNA, Complementary, Gene Library, Glutathione Transferase, Guanine Nucleotide Dissociation Inhibitors, Humans, Leukocytes, Lipid Metabolism, Molecular Sequence Data, Mutation, Protein Binding, Protein Prenylation, Protein Structure, Tertiary, Recombinant Fusion Proteins, Sequence Homology, Amino Acid, Two-Hybrid System Techniques, beta-Galactosidase, rho GTP-Binding Proteins
Amino Acid Sequence, DNA, Complementary, Gene Library, Glutathione Transferase, Guanine Nucleotide Dissociation Inhibitors, Humans, Leukocytes, Lipid Metabolism, Molecular Sequence Data, Mutation, Protein Binding, Protein Prenylation, Protein Structure, Tertiary, Recombinant Fusion Proteins, Sequence Homology, Amino Acid, Two-Hybrid System Techniques, beta-Galactosidase, rho GTP-Binding Proteins
Biochimie
Date: May. 01, 2001
PubMed ID: 11368848
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