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PCR cloning and expression analysis of cDNAs encoding cysteine proteinases from germinating seeds of Vicia sativa L.

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Abstract

cDNA clones encoding cysteine proteinases from cotyledons of germinated seeds of Vicia sativa L. have been obtained by means of PCR. Degenerate oligonucleotide primers were designed according to conserved amino acid regions of known cysteine proteinases. The deduced amino acid sequences of the cDNA clones encoding VSCYSPR1 and VSCYSPR2 display strong homology to cysteine proteinases of the so called papain superfamily. Northern analyses revealed developmentally regulated expression of both the mRNAs in germinating seeds. The transcripts were shown to be products of two distinct single genes, each exhibiting structural polymorphisms as exposed in few nucleotide substitutions.

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References

  1. Abe Y, Shirane K, Yokosawa H, Matsushita H, Mitta M, Kato I, Ishii S: Asparaginyl endopeptidase of jack bean seeds. J Biol Chem 268: 3525–3529 (1993).

    PubMed  Google Scholar 

  2. Baumgartner B, Chrispeels MJ: Purification and characterization of vicilin peptidohydrolase, the major endopeptidase in the cotyledons of mung bean seedlings. Eur J Biochem 77: 223–233 (1977).

    PubMed  Google Scholar 

  3. Cercos M, Carbonell J: Purification and characterization of a thiol-protease induced during senescence of unpollinated ovaries of Pisum sativum. Physiol Plant 88: 267–274 (1993).

    Article  Google Scholar 

  4. Granell A, Harris N, Pisabarro AG, Carbonell J: Temporal and spatial expression of a thiolprotease gene during pea ovary senescence, and its regulation by gibberellin. Plant J 2: 907–915 (1992).

    Article  PubMed  Google Scholar 

  5. Csoma C, Polgar L: Proteinase from germinating bean cotyledons. Biochem J 222: 769–776 (1984).

    PubMed  Google Scholar 

  6. Guerrero FD, Jones JT, Mullet JE: Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes. Plant Mol Biol 15: 11–26 (1990).

    PubMed  Google Scholar 

  7. Karrer KM, Pfeiffer SL, DiTomas ME: Two distinct gene subfamilies within the family of cysteine protease genes. Proc Natl Acad Sci USA 90: 3063–3067 (1993).

    PubMed  Google Scholar 

  8. Koehler SM, Ho THD: Hormonal regulation, processing and secretion of cysteine proteinases in barley aleurone layers. Plant Cell 2: 769–783 (1990).

    Article  PubMed  Google Scholar 

  9. Koizumi M, Yamaguchi-Shinozaki K, Tsuji H, Shinozaki K: Structure and expression of two genes that encode distinct drought-inducible cysteine proteinases in Arabidopsis thaliana. Gene 129: 175–182 (1993).

    Article  PubMed  Google Scholar 

  10. Lin E, Burns DJW, Gardner RC: Fruit developmental regulation of the kiwifruit actinidin promoter is conserved in transgenic petunia plants. Plant Mol Biol 23: 489–499 (1993).

    Article  PubMed  Google Scholar 

  11. Miller SH, Dykes DD, Polesky HF: A simple salting out procedure for extracting DNA from human nucleated cells. Nucl Acids Res 16: 1215 (1988).

    PubMed  Google Scholar 

  12. Mitsuhasi W, Koshiba T, Minamikawa T: Separation and characterization of two endopeptidases from cotyledons of germinating Vigna mungo seeds. Plant Physiol 80: 628–634 (1986).

    Google Scholar 

  13. Müntz K, Bassüner R, Lichtenfeld C, Scholz G, Weber E: Proteolytic cleavage of storage proteins during embryogenesis and germination of legume seeds. Physiol Vég 23: 75–79 (1985).

    Google Scholar 

  14. Muramatsu M, Fukazawa C: A high-order structure of plant storage proprotein allows its second conversion by an asparagine-specific cysteine protease, a novel proteolytic enzyme. Eur J Biochem 215: 123–132 (1993).

    PubMed  Google Scholar 

  15. Hara-Nishimura I, Inoue K, Nishimura M: A unique vacuolar processing enzyme responsible for conversion of several proprotein precursors into the mature forms. FEBS Lett 294: 89–93 (1991).

    Article  PubMed  Google Scholar 

  16. Hara-Nishimura I, Takeuchi Y, Nishimura M: Molecular characterization of a vacuolar processing enzyme related to a putative cysteine proteinase of Schistosoma mansoni. Plant Cell 5: 1651–1659 (1993).

    PubMed  Google Scholar 

  17. Schaffer MA, Fischer RL: Transcriptional activation by heat and cold of a thiol protease gene in tomato. Plant Physiol 93: 1486–1491 (1990).

    Google Scholar 

  18. Scott MP, Jung R, Müntz K, Nielsen NC: A protease responsible for post-translational cleavage of a conserved Asn-Gly linkage in glycinin, the major seed storage protein of soybean. Proc Natl Acad Sci USA 89: 658–662 (1992).

    PubMed  Google Scholar 

  19. Shutov AD, Lanh DN, Vaintraub IA: Isolation and partial characterization of protease B from germinating vetch seeds. Biokhimiya 47: 814–821 (1982).

    Google Scholar 

  20. Shutov AD, Vaintraub IA: Degradation of storage proteins in germinating seeds. Phytochemistry 26: 1557–1566 (1987).

    Article  Google Scholar 

  21. Tanaka T, Yamauchi D, Minamikawa T: Nucleotide sequence of cDNA for an endopeptidase (EP-C1) from pods of maturing Phaseolus vulgaris fruits. Plant Mol Biol 16: 1083–1084 (1991).

    Google Scholar 

  22. Watanabe H, Abe K, Emori Y, Hosoyama H, Arai S: Molecular cloning and gibberellin-induced expression of multiple cysteine proteinases of rice seeds (oryzains). J Biol Chem 266: 16897–16902 (1991).

    PubMed  Google Scholar 

  23. Yamauchi D, Akasofu H, Minamikawa T: Cysteine endopeptidase from Vigna mungo: gene structure and expression. Plant Cell Physiol 33: 789–797 (1992).

    Google Scholar 

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Becker, C., Fischer, J., Nong, V.H. et al. PCR cloning and expression analysis of cDNAs encoding cysteine proteinases from germinating seeds of Vicia sativa L.. Plant Mol Biol 26, 1207–1212 (1994). https://doi.org/10.1007/BF00040701

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  • DOI: https://doi.org/10.1007/BF00040701

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