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A novel ras-related rgp1 gene encoding a GTP-binding protein has reduced expression in 5-azacytidine-induced dwarf rice

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Summary

Exposure of normal, tall rice (Oryza sativa) seedlings to 5-azacytidine, a powerful inhibitor of DNA methylation in vivo, induced both demethylation of genomic DNA and dwarf plants. Genes that had been affected by treatment were identified by differential screening of a cDNA library, and a ras-related gene, rgp1, was subsequently isolated. The cDNA of rgp1 was found to encode a deduced protein sequence of 226 amino acids with a relative molecular mass of 24850, which was most closely related to the ras-related ypt3 protein of fission yeast, Shizosaccharomyces pombe. The rgp1 protein, expressed in transformed Escherichia coli, clearly showed GTP-binding activity. During seedling growth, rgp1 expression was first observed 14 days after germination, reaching a maximum level between 28 and 42 days, and gradually decreased thereafter until 63 days when it attained the same level of expression as in 14-day-old seedlings. Expression of rgp1 was found to be markedly reduced throughout the growth period of both 5-azacytidine-induced dwarf plants and their progenies, relative to levels in untreated tall control plants. These results suggest that expression of rgp1 may be influenced, either directly or indirectly, by DNA methylation, and that the rgp1 protein may play an important role in plant growth and development.

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References

  • Adams RLP, Burdon RH (1985) Molecular biology of DNA methylation. Springer-Verlag, New York

    Google Scholar 

  • Balch WE (1989) Biochemistry of interorganelle transport. J Biol Chem 264:16965–16968

    Google Scholar 

  • Barbacid M (1987) ras genes. Annu Rev Biochem 56:779–827

    Google Scholar 

  • Bourne HR (1988) Do GTPases direct membrane traffic in secretion? Cell 53:669–671

    Google Scholar 

  • Bucci C, Frunzio R, Chiariotti L, Brown AL, Rechler MM, Brum CB (1988) A new member of the ras gene superfamily identified in a rat liver cell line. Nucleic Acids Res 16:9979–9986

    Google Scholar 

  • Cedar H, Razin A (1990) DNA methylation and development. Biochim Biophys Acta 1049:1–8

    Google Scholar 

  • Chardin P, Tavitian A (1986) The ral gene: A new ras related gene isolated by the use of a synthetic probe. EMBO J 5:2203–2208

    Google Scholar 

  • Chardin P, Tavitian A (1989) Coding sequence of human ralA and ralB cDNAs. Nucleic Acids Res 17:4380

    Google Scholar 

  • Chavrier P, Parton RG, Hauri HP, Simons K, Zerial M (1990) Localization of low molecular weight GTP binding proteins to exocytic and endocytic compartments. Cell 62:317–329

    Google Scholar 

  • Darchen F, Zahraoui A, Hammel F, Monteils M-P, Tavitian A, Scherman D (1990) Association of the GTP-binding protein Rab3A with bovine adrenal chromaffin granules. Proc Natl Acad Sci USA 87:5692–5696

    Google Scholar 

  • de Vos AM, Tong L, Milburn MV, Matias PM, Jancarik J, Noguchi S, Nishimura S, Miura K, Ohtsuka E, Kim S-H (1988) Three-dimensional structure of an oncogene protein: catalytic domain of human c-H-ras-p21. Science 239:888–893

    Google Scholar 

  • Drivas GT, Shih A, Coutavas E, Rush MG, D'eustachio P (1990) Characterization of four novel ras-like genes expressed in a human teratocarcinoma cell line. Mol Cell Biol 10:1793–1798

    Google Scholar 

  • Gallwitz D, Donath C, Sander C (1983) A yeast gene encoding a protein homologous to the human c-has/bas proto-oncogene product. Nature 306:704–707

    Google Scholar 

  • Goud B, Zahraoui A, Tavitian A, Saraste J (1990) Small GTP-binding protein associated with Golgi cisternae. Nature 345:553–556

    Google Scholar 

  • Gruenbaum Y, Naveh-Many T, Cedar H, Razin A (1981) Sequence specificity of methylation in higher plant DNA. Nature 292:860–862

    Google Scholar 

  • Hall A (1990) ras and GAP — who's controlling whom? Cell 61:921–923

    Google Scholar 

  • Haubruck H, Engelke U, Mertins P, Gallwitz D (1990) Structural and functional analysis of ypt2, and essential ras-related gene in the fission yeast Schizosaccharomyces pombe encoding a Sec4 protein homologue. EMBO J 9:1957–1962

    Google Scholar 

  • Hengst L, Lehmeier T, Gallwitz D (1990) The ryh1 gene in the fission yeast Schizosaccharomyces pombe encoding a GTP-binding protein related to ras, rho and ypt: structure, expression and identification of its human homologue. EMBO J 9:1949–1955

    Google Scholar 

  • Holliday R (1987) The inheritance of epigenetic defects. Science 238:163–170

    Google Scholar 

  • Klickstein LB, Neve RL (1987) Preparation of insert DNA from messenger RNA. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current protocols in molecular biology. Wiley, New York, pp 5.5–5.6

    Google Scholar 

  • Madaule P, Axel R, Myers AM (1987) Characterization of two members of the rho gene family from the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci USA 84:779–783

    Google Scholar 

  • Maniatis T, Fritsch ET, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Matsui M, Sasamoto S, Kunieda T, Nomura N, Ishizaki R (1989) Cloning of ara, a putative Arabidopsis thaliana gene homologous to the ras-related gene family. Gene 76:331–319

    Google Scholar 

  • McCormick F (1989) ras GTPase activating protein: signal transmitter and signal terminator. Cell 56:5–8

    Google Scholar 

  • Mitchell PJ, Tjian R (1989) Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science 245:371–378

    Google Scholar 

  • Miyake S, Yamamoto M (1990) Identification of ras-related, YPT-family genes in Schizosaccharomyces pombe. EMBO J 9:1417–1422

    Google Scholar 

  • Molenaar CMT, Prange R, Gallwitz D (1988) A carboxyl-terminal cysteine residue is required for palmitic acid binding and biological activity of the ras-related yeast YPT1 protein. EMBO J 7:971–976

    Google Scholar 

  • Morre DJ, Mollenhauer HH (1976) Interactions among cytoplasm, endomembranes and the cell surface. In: Stocking CR, Heber U (eds) Transport in plants III. Springer Verlag, New York, pp 288–344

    Google Scholar 

  • Nick H, Brwen B, Ferl RJ, Gilbert W (1986) Detection of cytosine methylation in the maize alcohol dehydrogenase gene by genomic sequencing. Nature 319:243–246

    Google Scholar 

  • Pai EF, Kabsch W, Krengel U, Holmes KC, John J, Wittinghofer A (1989) Structure of the guanine-nucleotide-binding domain of the Ha-ras oncogene product p21 in the triphosphate conformation. Nature 341:209–214

    Google Scholar 

  • Pizon V, Lerosey I, Chardin P, Tavitian A (1988) Nucleotide sequence of a human cDNA encoding a ras-related protein (rap1B). Nucleic Acids Res 16:7719

    Google Scholar 

  • Razin A, Cedar H, Riggs AD (1984) DNA methylation. Biochemistry and biological significance. Springer-Verlag, New York

    Google Scholar 

  • Reece KS, McElroy D, Wu R (1990) Genomic nucleotide sequence of four rice (Oryza sativa) actin genes. Plant Mol Biol 14:621–624

    Google Scholar 

  • Sano H, Kamada I, Youssefian S, Katsumi M, Wabiko H (1990) A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol Gen Genet 220:441–447

    Google Scholar 

  • Schmitt HD, Wagner P, Pfaff E, Gallwitz D (1986) The ras-related YPT1 gene product in yeast: A GTP-binding protein that might be involved in microtubule organization. Cell 47:401–412

    Google Scholar 

  • Segev N, Mulholland J, Botstein D (1988) The yeast GTP-binding YPT1 protein and a mammalian counterpart are associated with the secretion machinery. Cell 52:915–924

    Google Scholar 

  • Sigal IS, Gibbs JB, D'Alonzo JS, Temeles GL, Wolanski BS, Socher SH, Scolnick EM (1986) Mutant ras-encoded proteins with altered nucleotide binding exert dominant biological effects. Proc Natl Acad Sci USA 83:952–956

    Google Scholar 

  • Touchot N, Chardin P, Tavitian A (1987) Four additional members of the ras gene superfamily isolated by an oligonucleotide strategy: Molecular cloning of YPT-related cDNAs from a rat brain library. Proc Natl Acad Sci USA 84:8210–8214

    Google Scholar 

  • Walworth N, Goud B, Kabcenell A, Novick P (1989) Mutational analysis of SEC4 suggests a cyclical mechanism for the regulation of vesicular traffic. EMBO J 8:1685–1693

    Google Scholar 

  • Zabulionis RB, Atkinson BG, Procunier JD, Walden DB (1988) Maize (Zea mays L.) DNA sequences homologous to the oncegenes myb, ras, and src. Genome 30:820–824

    Google Scholar 

  • Zahraoui A, Touchot N, Chardin P, Tavitian A (1989) The human Rab genes encode a family of GTP-binding proteins related to yeast YPT1 and SEC4 products involved in secretion. J Biol Chem 264:12394–12401

    Google Scholar 

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Communicated by H. Saedler

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Sano, H., Youssefian, S. A novel ras-related rgp1 gene encoding a GTP-binding protein has reduced expression in 5-azacytidine-induced dwarf rice. Molec. Gen. Genet. 228, 227–232 (1991). https://doi.org/10.1007/BF00282470

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