Abstract
We report here the cloning and sequence analysis of cDNAs for a pair of closely related proteins from soybean (Glycine max [L.] Merr. cv. Williams 82) stems. Both proteins are abundant in soluble extracts of seedling stems but not of roots. One of these proteins (M r=28 kDa) is also foundd in the cell wall fraction of stems and actumulates there when seedlings are exposed to mild water deficit for 48 h. The mRNA for these proteins is most abundant in the stem region which contains dividing cells, less abundant in elongating and mature stem cells, and rare in roots. Using antiserum against the 28 kDa protein, we isolated cDNA clones encoding it and an antigenically related 31 kDa protein. The two cDNAs are 80% homologous in nucleotide and amino acid coding sequence. The predicted proteins have similar hydropathy profiles, and contain putative NH2-terminal signal sequences and a single putative N-linked glycosylation site. The two proteins differ significantly in calculated pI (28 kDa=8.6; 31 kDa=5.8), and the charge difference is demonstrated on two-dimensional gels. The proteins described here may function as somatic storage proteins during early seedling development, and are closely related to glycoproteins which accumulate in vacuoles of paraveinal mesophyll cells of fully expanded soybean leaves when plants are depodded.
Similar content being viewed by others
References
Bantle JA, Maxwell IA, Hahn WE: Specificity of oligo(dT)-cellulose chromatography in the isolation of polyadenylated RNA. Anal Biochem 72: 413–427 (1976).
Bard J, Bourque DP, Hildebrand M, Zaitlin D: In vitro expression of chloroplast genes in lysates of higher plant chloroplasts. Proc Natl Acad Sci USA 82: 3983–3987 (1985).
Beevers L: Post-translational modifications. In: Boulter D, Parthier B (eds) Encyclopedia of Plant Physiology, New Series Vol. 14A, pp. 136–168. Springer-Verlag, Berlin (1982).
Bensen RJ, Boyer JS, Mullet JE: Water deficit induced changes in abscisic acid, growth, polysomes and translatable RNA in soybean stems. Plant Physiol, in press (1988).
Bozarth CS, Mullet JE, Boyer JS: Cell wall proteins at low water potentials. Plant Physiol 85: 261–267 (1987).
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).
Breitbart RE, Andreadis A, Nadal-Ginard B: Alternative splicing: a ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Ann Rev Biochem 56: 467–495 (1987).
Cavlieri AJ, Boyer JS: Water potentials induced by growth in soybean hypocotyls. Plant Physiol 69: 492–496 (1982).
Franceschi VR, Giaquinta RT: The paraveinal mesophyll of soybean leaves in relation to assimilate transfer and compartmentation. II. Structural, metabolic and compartmental changes during reproductive growth. Planta 157: 422–431 (1983).
Franceschi VR, Wittenbach VA, Giaquinta RT: Paraveinal mesophyll of soybean leaves in relation to assimilate transfer and compartmentation. III. Immunohistochemical localization of specific glycopeptides in the vacuole after depodding. Plant Physiol 72: 586–589 (1983).
Gasser SM, Ohashi A, Daum G, Bohni PC, Gibson J, Reid GA, Yonetani T, Schatz G: Imported mitochondrial proteins cytochrome b 2 and cytochrome c 1 are processed in two steps. Proc Natl Acad Sci USA 79: 267–271 (1982).
Gubler U, Hoffman BJ: A simple and very efficient method for generating cDNA libraries. Gene 25: 263–269 (1983).
Hopp TP, Woods KR: Prediction of antigenic determinants from amino acid sequences. Proc Natl Acad Sci USA 78: 3824–3828 (1981).
Huynh TV, Young RA, Davis RW: Construction and screening of cDNA libraries in λgt10 and λgt11. In: Glover D (ed) DNA Cloning Techniques: a Practical Approach, pp 49–78. IRL Press, Oxford (1985).
Ikeuchi M, Plumley FG, Inoue Y, Schmidt GW: Phosphorylation of photosystem II components. CP43 apoprotein, D1, D2, and 10 to 11 kilodalton protein in chloroplast thylakoids of higher plants. Plant Physiol 85: 638–642 (1987).
Johnson LM, Bankaitis VA, Emr SD: Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease. Cell 48: 875–885 (1987).
Kornfeld R, Kornfeld S: Assembly of asparagine-linked oligosaccharides. Ann Rev Biochem 54: 631–644 (1985).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Mason HS, Mullet JE, Boyer JS: Polysomes, messenger RNA and growth in soybean stems during development and water deficit. Plant Physiol 86: 725–733 (1988).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Meyer RF, Boyer JS: Osmoregulation, solute distribution, and growth in soybean seedlings having low water potentials. Planta 151: 482–489 (1981).
Mishkind ML, Wessler SR, Schmidt GW: Functional determinants in transit sequences: import and partial maturation by vascular plant chloroplasts of the ribulose-1,5-bisphospate carboxylase small subunit of Chlamydomonas. J Cell Biol 100: 226–234 (1985).
O'Farrell PH: High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021 (1975).
O'Farrell PZ, Goodman HM, O'Farrell PH: High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12: 1133–1142 (1977).
Paiva E, Lister RM, Park WD: Induction and accumulation of major tuber proteins of potato in stems and petioles. Plant Physiol 71: 161–168 (1983).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Singh NK, Bracker CA, Hasegawa PM, Handa AK, Buckel S, Hermodson MA, Pfankoch E, Regnier FE, Bressan RA: Characterization of osmotin, a thaumatin-like protein associated with osmotic adaptation in plant cells. Plant Physiol 85: 529–536 (1987).
Staswick P: Soybean vegetative storage protein structure and gene expression. Plant Physiol 87: 250–254 (1988).
Tague BW, Chrispeels MJ: The plant vacuolar protein, phytohemagglutinin, is transported to the vacuole of transgenic yeast. J Cell Biol 105: 1971–1979 (1987).
Tague BW, Chrispeels MJ: Identification of a plant vacuolar targeting signal in yeast. Plant Physiol 86: 84a (1988).
Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354 (1979).
VonHeijne G: A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 14: 4683–4690 (1986).
Wittenbach VA: Purification and characterization of a soybean leaf storage glycoprotein. Plant Physiol 73: 125–129 (1983).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mason, H.S., Guerrero, F.D., Boyer, J.S. et al. Proteins homologous to leaf glycoproteins are abundant in stems of dark-grown soybean seedlings. Analysis of proteins and cDNAs. Plant Mol Biol 11, 845–856 (1988). https://doi.org/10.1007/BF00019524
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00019524