Rom1p and Rom2p are GDP/GTP exchange proteins (GEPs) for the Rho1p small GTP binding protein in Saccharomyces cerevisiae.

The RHO1 gene encodes a homolog of the mammalian RhoA small GTP binding protein in the yeast Saccharomyces cerevisiae. Rho1p is localized at the growth site and is required for bud formation. Multicopy suppressors of a temperature‐sensitive, dominant negative mutant allele of RHO1, RHO1(G22S, D125N), were isolated and named ROM (RHO1 multicopy suppressor). Rom1p and Rom2p were found to contain a DH (Dbl homologous) domain and a PH (pleckstrin homologous) domain, both of which are conserved among the GDP/GTP exchange proteins (GEPs) for the Rho family small GTP binding proteins. Disruption of ROM2 resulted in a temperature‐sensitive growth phenotype, whereas disruption of both ROM1 and ROM2 resulted in lethality. The phenotypes of deltarom1deltarom2 cells were similar to those of deltarho1 cells, including growth arrest with a small bud and cell lysis. Moreover, the temperature‐sensitive growth phenotype of deltarom2 was suppressed by overexpression of RHO1 or RHO2, but not of CDC42. The glutathione‐S‐transferase (GST) fusion protein containing the DH domain of Rom2p showed the lipid‐modified Rholp‐specific GDP/GTP exchange activity which was sensitive to Rho GDP dissociation inhibitor. These results indicate that Rom1p and Rom2p are GEPs that activate Rho1p in S.cerevisiae.

[1]  C. Tognon,et al.  Expression Cloning of lfc, a Novel Oncogene with Structural Similarities to Guanine Nucleotide Exchange Factors and to the Regulatory Region of Protein Kinase C (*) , 1995, The Journal of Biological Chemistry.

[2]  T. Sasaki,et al.  Rho as a regulator of the cytoskeleton. , 1995, Trends in biochemical sciences.

[3]  John G. Collard,et al.  A role for Rac in Tiaml-induced membrane ruffling and invasion , 1995, Nature.

[4]  M. Olson,et al.  Direct Involvement of the Small GTP-binding Protein Rho in lbc Oncogene Function (*) , 1995, The Journal of Biological Chemistry.

[5]  J. Chant,et al.  GTPase cascades choreographing cellular behavior: Movement, morphogenesis, and more , 1995, Cell.

[6]  G M Bokoch,et al.  Guanine nucleotide exchange regulates membrane translocation of Rac/Rho GTP-binding proteins. , 1994, The Journal of biological chemistry.

[7]  R. Cerione,et al.  Interactions between the bud emergence proteins Bem1p and Bem2p and Rho- type GTPases in yeast , 1994, The Journal of cell biology.

[8]  K. Sakaguchi,et al.  A novel oncogene, ost, encodes a guanine nucleotide exchange factor that potentially links Rho and Rac signaling pathways. , 1994, The EMBO journal.

[9]  P. Hajduk,et al.  Pleckstrin homology domains bind to phosphatidylinositol-4,5-bisphosphate , 1994, Nature.

[10]  S. Powers,et al.  Ras-15A protein shares highly similar dominant-negative biological properties with Ras-17N and forms a stable, guanine-nucleotide resistant complex with CDC25 exchange factor. , 1994, Oncogene.

[11]  T J Gibson,et al.  PH domain: the first anniversary. , 1994, Trends in biochemical sciences.

[12]  K. Tanaka,et al.  Molecular cloning and characterization of yeast rho GDP dissociation inhibitor. , 1994, The Journal of biological chemistry.

[13]  K. Tanaka,et al.  Growth site localization of Rho1 small GTP-binding protein and its involvement in bud formation in Saccharomyces cerevisiae , 1994, The Journal of cell biology.

[14]  John G. Collard,et al.  Identification of an invasion-inducing gene, Tiam-1, that encodes a protein with homology to GDP-GTP exchangers for Rho-like proteins , 1994, Cell.

[15]  R. Lefkowitz,et al.  Binding of G protein beta gamma-subunits to pleckstrin homology domains. , 1994, The Journal of biological chemistry.

[16]  T. Fleming,et al.  Expression cDNA cloning of a novel oncogene with sequence similarity to regulators of small GTP-binding proteins. , 1994, Oncogene.

[17]  David A. Williams,et al.  Novel human oncogene lbc detected by transfection with distinct homology regions to signal transduction products. , 1994, Oncogene.

[18]  T. Sasaki,et al.  The Dbl oncogene product as a GDP/GTP exchange protein for the Rho family: its properties in comparison with those of Smg GDS. , 1994, Biochemical and biophysical research communications.

[19]  Y. Zheng,et al.  Control of the yeast bud-site assembly GTPase Cdc42. Catalysis of guanine nucleotide exchange by Cdc24 and stimulation of GTPase activity by Bem3. , 1994, The Journal of biological chemistry.

[20]  S. Aaronson,et al.  Cellular transformation and guanine nucleotide exchange activity are catalyzed by a common domain on the dbl oncogene product. , 1994, The Journal of biological chemistry.

[21]  A. Hall,et al.  Small GTP-binding proteins and the regulation of the actin cytoskeleton. , 1994, Annual review of cell biology.

[22]  Mark S. Boguski,et al.  Proteins regulating Ras and its relatives , 1993, Nature.

[23]  J. Thompson,et al.  The PH domain: a common piece in the structural patchwork of signalling proteins. , 1993, Trends in biochemical sciences.

[24]  V. Kaartinen,et al.  Human ABR encodes a protein with GAPrac activity and homology to the DBL nucleotide exchange factor domain. , 1993, The Journal of biological chemistry.

[25]  L. Staudt,et al.  Ly-GDI, a GDP-dissociation inhibitor of the RhoA GTP-binding protein, is expressed preferentially in lymphocytes. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Jonathan A. Cooper,et al.  Mammalian Ras interacts directly with the serine/threonine kinase raf , 1993, Cell.

[27]  S. Fields,et al.  Elimination of false positives that arise in using the two-hybrid system. , 1993, BioTechniques.

[28]  M. Boguski,et al.  Influence of guanine nucleotides on complex formation between Ras and CDC25 proteins , 1993, Molecular and cellular biology.

[29]  J. Lélias,et al.  cDNA cloning of a human mRNA preferentially expressed in hematopoietic cells and with homology to a GDP-dissociation inhibitor for the rho GTP-binding proteins. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Masato Ishikawa,et al.  Constraint Based Alignment Editor with Automatic Alinger and Motif Dictionary Retriever , 1993 .

[31]  T. Sasaki,et al.  Post-translational processing of rac p21s is important both for their interaction with the GDP/GTP exchange proteins and for their activation of NADPH oxidase. , 1992, The Journal of biological chemistry.

[32]  J. Carpentier,et al.  The osmotic integrity of the yeast cell requires a functional PKC1 gene product , 1992, Molecular and cellular biology.

[33]  C. Wollheim,et al.  The small GTP-binding proteins in the cytosol of insulin-secreting cells are complexed to GDP dissociation inhibitor proteins. , 1992, The Journal of biological chemistry.

[34]  T. Sasaki,et al.  Functional interactions of stimulatory and inhibitory GDP/GTP exchange proteins and their common substrate small GTP-binding protein. , 1992, The Journal of biological chemistry.

[35]  E. Martegani,et al.  Cloning by functional complementation of a mouse cDNA encoding a homologue of CDC25, a Saccharomyces cerevisiae RAS activator. , 1992, The EMBO journal.

[36]  T. Sasaki,et al.  Cooperative function of rho GDS and rho GDI to regulate rho p21 activation in smooth muscle. , 1992, Biochemical and biophysical research communications.

[37]  R. Schiestl,et al.  Improved method for high efficiency transformation of intact yeast cells. , 1992, Nucleic acids research.

[38]  S. Aaronson,et al.  Catalysis of guanine nucleotide exchange on the CDC42Hs protein by the dbloncogene product , 1991, Nature.

[39]  K. Kaibuchi,et al.  A stimulatory GDP/GTP exchange protein for smg p21 is active on the post-translationally processed form of c-Ki-ras p21 and rhoA p21. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[40]  D. Drubin Development of cell polarity in budding yeast , 1991, Cell.

[41]  K. Kaibuchi,et al.  Molecular cloning of the cDNA for stimulatory GDP/GTP exchange protein for smg p21s (ras p21-like small GTP-binding proteins) and characterization of stimulatory GDP/GTP exchange protein. , 1991, Molecular and cellular biology.

[42]  J. Cleveland,et al.  Loss of the amino-terminal helix-loop-helix domain of the vav proto-oncogene activates its transforming potential , 1991, Molecular and cellular biology.

[43]  Frank McCormick,et al.  The GTPase superfamily: conserved structure and molecular mechanism , 1991, Nature.

[44]  J. Pringle,et al.  CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae , 1990, The Journal of cell biology.

[45]  J. Pringle,et al.  Molecular characterization of CDC42, a Saccharomyces cerevisiae gene involved in the development of cell polarity , 1990, The Journal of cell biology.

[46]  S. Aaronson,et al.  The N-terminal region of proto-dbl down regulates its transforming activity. , 1989, Oncogene.

[47]  S. Fields,et al.  A novel genetic system to detect protein–protein interactions , 1989, Nature.

[48]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[49]  M. Wigler,et al.  Dominant yeast and mammalian RAS mutants that interfere with the CDC25-dependent activation of wild-type RAS in Saccharomyces cerevisiae , 1989, Molecular and cellular biology.

[50]  R. Higuchi Using PCR to Engineer DNA , 1989 .

[51]  K. Ikeda,et al.  Purification and characterization of a novel GTP-binding protein with a molecular weight of 24,000 from bovine brain membranes. , 1988, The Journal of biological chemistry.

[52]  Y. Matsuura,et al.  Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. , 1987, The Journal of general virology.

[53]  G. Fink,et al.  KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast , 1987, Cell.

[54]  R. Axel,et al.  Characterization of two members of the rho gene family from the yeast Saccharomyces cerevisiae. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[55]  A. Myers,et al.  Yeast/E. coli shuttle vectors with multiple unique restriction sites , 1986, Yeast.

[56]  S. Henikoff Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. , 1984, Gene.

[57]  L. Guarente Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. , 1983, Methods in enzymology.

[58]  D. Botstein,et al.  Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase , 1982, Cell.

[59]  K. Kaibuchi,et al.  A stimulatory GDP / GTP exchange protein for smg p 21 is active on the post-translationally processed form of cKi-ras p 21 and rhoA p 21 ( ras p 21 / rho p 21 / prenylation ) , 2022 .