Abstract
Developmentally regulated GTP-binding proteins (DRGs) from animals and fungi are highly conserved but have no known function. Here we characterize DRGs from pea (PsDRG) and Arabidopsis (AtDRG). Amino acid sequences of AtDRG and PsDRG were 90% identical to each other and about 65% identical to human DRG. Genomic Southern blotting indicated that AtDRG and PsDRG probably are single-copy genes. PsDRG mRNA accumulated preferentially in growing organs (root apices, growing axillary buds and elongating stems) compared with their non-growing counterparts. At DRG mRNA was relatively abundant in Arabidopsis leaves, stems and siliques, less abundant in flowers and flower buds, and barely detectable in roots. Histone mRNAs are known to accumulate predominantly during S phase of the cell cycle and are markers for proliferating cells. The patterns of histone H2A mRNA accumulation in pea and Arabidopsis organs were very similar to those of DRG mRNAs. An antiserum raised against a PsDRG N-terminal fusion protein recognized 43 and 45 kDa proteins. PsDRG proteins were more abundant in growing pea roots and stems than in non-growing organs, but they were equally abundant in growing and dormant axillary buds. After differential centrifugation, PsDRG proteins were found primarily in the microsomal (150 000×g pellet) and soluble (150 000×g supernatant) cell fractions.
Similar content being viewed by others
References
Barbacid M: ras genes. Annu Rev Biochem 56: 779–827 (1987).
Borg S, Brandstrup B, Jensen TJ, Poulen C: Identification of a new protein species among 33 different small GTP-binding proteins encoded by cDNAs from Lotus japonicus, and expressing of corresponding mRNAs in developing root nodules. Plant J 11: 237–250 (1997).
Bourne HR, Sanders DA, McCormick F: The GTPase superfamily: a conserved switch for diverse cell functions. Nature 349: 117–127 (1990).
Bourne HR, Sanders DA, McCormick F: The GTPase superfamily: conserved structure and molecular mechanism. Nature 349: 117–127 (1991).
Bussey H, Kaback DB, ZhongWW, Vo DT, Clark MW, Fortin N, Hall J, Ouellette BFF, Keng T, Barton AB, Su Y, Davies CJ, Storms RK: The nucleotide sequence of chromosome I from Saccharomyces cerevisiae. Proc Natl Acad Sci USA 92: 3809–3813 (1995).
Clemons DJ, Besch-Williford C, Steffen EK, Riley LK, Moore DH: Evaluation of a subcutaneously implanted chamber for antibody production in rabbits. Lab Animal Sci 42: 307–311 (1992).
Devitt ML, Stafstrom JP: Cell cycle regulation during growthdormancy cycles in pea axillary buds. Plant Mol Biol 29: 255–265 (1995).
Dombrowski JE, Raikhel NV: Isolation of a cDNA encoding a novel GTP-binding protein of Arabidopsis thaliana. PlantMol Biol 28: 1121–1126 (1995).
Gilman AG: G-proteins: transducers of receptor-generated signals. Annu Rev Biochem 56: 615–649 (1987).
Glomset JA, Farnsworth CC: role of protein modification in programming interactions between ras-related GTPases and cell membranes. Annu Rev Cell Biol 10: 181–205 (1994).
Gu X, Verma DP: Phragmoplastin, a dynamin-like protein associated with cell plate formation in plants. EMBO J 15: 695–704 (1996).
Hall A: Small GTP-binding proteins and the regulation of the actin cytoskeleton. Annu Rev Cell Biol 10: 31–54 (1994).
Hudson JD, Young PG: Sequence of the Schizosaccharomyces pombe gtp1 gene and identification of a novel family of putative GTP-binding proteins. Gene 125: 191–193 (1993).
Hwang YW, Sanchez A, Miller DL: Mutagenesis of bacterial elongation factor Tu at lysine 136. A conserved amino acid in GTP regulatory proteins. J Biol Chem 264: 8304–8309 (1989).
Ishida S, Takahashi Y, Nagata T: The mode of expression and promoter analysis of the arcA gene, an auxin-regulated gene in tobacco BY-2 cells. Plant Cell Physiol 37: 439–448 (1996).
Johnson DR, Bhatnagar RS, Knoll LJ, Gordon JI: Genetic and biochemical studies of protein N-myristoylation. Annu Rev Biochem 63: 869–914 (1994).
Kumar S, Tomooka Y, Noda M: Identification of a set of genes with developmentally down-regulated expression in the mouse brain. Biochem Biophys Res Comm 185: 1155–1161 (1992).
Kumar S, Iwao M, Yamagishi T, Noda M, Asahima M: Expression of GTP-binding protein gene drg during Xenopus laevis development. Int J Dev Biol 37: 539–546 (1993).
Ma H, Yanofsky MF, Meyerowitz EM: Molecular cloning and characterization of GPA1, a G protein_ subunit gene from Arabidopsis thaliana. Proc Natl Acad Sci USA 87: 3821–3825 (1990).
Mahajan MA, Part ST, Sun XH: Association of a novel GTP binding protein, DRG, with Tal1 oncogenic proteins. Oncogene 12: 2343–2350 (1996).
Moroianu J, Blobel G: Protein export from the nucleus requires the GTPase Ran and GTP hydrolis. Proc Natl Acad Sci USA 92: 4318–4322 (1995).
Moss J, Vaughan M: ADP-rybosylation factors, 20,000 Mr guanine nucleotide-binding protein activators of cholera toxin and components of intracellular vesicular transport systems. Cell Signal 5: 367–379 (1993).
Newman T, de Bruijn FJ, Green P, Keegstra K, Kende H, McIntosh L, Ohlrogge J, Raikhel N, Somerville S, Thomshow M, Retzel E, Somerville C: Genes galore: a summary of methods for accessing results from large-scale partial sequencing of anonymous Arabidopsis cDNA clones. Plant Physiol 106: 1241–1255 (1994).
Nuoffer C, Balch WE: GTPases: multifunctional molecular switches regulating vesicular traffic. Annu Rev Biochem 63: 949–990 (1994).
Rost R, Jones TJ, Falk RH: Distribution and relationship of cell division and maturation events in Pisum sativum (Fabaceae) seedling roots. Am J Bot 75: 1571–1583 (1988).
Sazuka T, Kinoshita M, Tomooka Y, Ikawa Y, Noda M, Kumar S: Expression of DRG during mouse embryonic development. Biochem Biophys Res Comm 189: 371–377 (1992).
Sazuka T, Tomooka Y, Ikawa Y, Noda M, Kumar S: DRG: a novel developmentally regulated GTP-binding protein. Biochem Biophys Res Comm 189: 363–370 (1992).
Schenker T, Lach C, Kessler B, Calderara S, Treub B: A novel GTP-binding protein which is selectively repressed in SV40 transformed fibroblasts. J Biol Chem 269: 25447–25453 (1994).
Shimmin LC, Dennis PP: Characterization of the L11, L1, L10, L12 equivalent ribosomal protein gene cluster of the halophilic archaebacterium Halobacterium cutirubrum. EMBO J 8: 1225–1235 (1989).
Sommer KA, Petersen G, Bautz EKF: The gene upstream of DmRP128 codes for a novel GTP-binding protein of Drosophila melanogaster. Mol Gen Genet 242: 391–398 (1994).
Staehelin LA, Moore I: The plant golgi apparatus: structure, functional organization and trafficking mechanisms. Annu Rev Plant Physiol Plant Mol Biol 46: 261–288 (1995).
Stafstrom JP, Ripley BD, Devitt ML, Drake B: Dormancyassociated gene expression in pea axillary buds. Cloning and expression of PsDRM1 and PsDRM2. Planta 205: 547–552 (1998).
Stafstrom JP, Sussex IM: Expression of a ribosomal protein gene in axillary buds of pea. Plant Physiol 100: 1494–1502 (1992).
Tanimoto EY, Rost TL, Comai L: DNA replication-dependent histone H2A mRNA expression in pea root tips. Plant Physiol 103: 1291–1297 (1993).
Terryn N, Van Montagu M, Inzé D: GTP-binding proteins in plants. Plant Mol Biol 22: 143–152 (1993).
Wilson R and 54 others: 2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans. Nature 368: 32–38 (1994).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Devitt, M.L., Maas, K.J. & Stafstrom, J.P. Characterization of DRGs, developmentally regulated GTP-binding proteins, from pea and Arabidopsis. Plant Mol Biol 39, 75–82 (1999). https://doi.org/10.1023/A:1006178710443
Issue Date:
DOI: https://doi.org/10.1023/A:1006178710443