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Transcript levels for a mung bean cysteine protease during early seedling growth

Published online by Cambridge University Press:  19 September 2008

Kangmoon Lee ,
Zhouwen Liu ,
Anna Tan-Wilson  and
Karl Wilson
Kangmoon Lee
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902-6000, USA
Zhouwen Liu
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902-6000, USA
Anna Tan-Wilson
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902-6000, USA
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Abstract

cDNAs for the major cysteine endopeptidase (CEpase) of mung bean (Vigna radiata [L.] Wilczek) seedling cotyledons have been cloned using gene-specific primers with the polymerase chain reaction (PCR) in a 3'-RACE system. A cDNA clone for CEpase, pKL042, that is 1221 bp long excluding the poly A tail was isolated. It appears to contain the entire coding sequence for a 362-residue-long polypeptide. The N-terminal sequence for the mature CEpase begins at position 128 of the putative translation product, suggesting removal of an N-terminal hydrophobic signal peptide and additional sequences to produce the mature protease. Northern blot hybridization with CEpase cDNA pKL042 as probe indicates that CEpase transcript is not detectable in the cotyledons or the embryonic axis of dry seeds, but is first detectable in the day 1 cotyledons and in the day 3 axis. The level of CEpase mRNA in both cotyledons and axis increases as growth proceeds. A decline of protease activity, however, is observed after day 3 in the cotyledons, even though the level of protease transcripts continues to increase until day 8. Detachment of the axis from the cotyledons before day 3 results in the prevention of the normal increase in both protease activity and CEpase mRNA.

Keywords

Cysteine endopeptidasecDNAexpressionregulationmung bean
Type
Physiology and Biochemistry
Information
Seed Science Research , Volume 7 , Issue 4 , December 1997 , pp. 359 - 372
Copyright
Copyright © Cambridge University Press 1997

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References

Akasofu, H., Yamauchi, D., Mitsuhashi, W. and Minamikawa, T. (1989) Nucleotide sequence of cDNA for sulfhydryl-endopeptidase (SH-EP) from cotyledons of germinating Vigna mungo seeds. Nucleic Acids Research 17, 6733.CrossRefGoogle ScholarPubMed
Baumgartner, B. and Chrispeels, M.J. (1977) Purification and characterization of vicilin peptidohydrolase, the major endopeptidase in the cotyledons of mung bean seedlings. European Journal of Biochemistry 77, 223–233.CrossRefGoogle ScholarPubMed
Baumgartner, B., Tokuyasu, K.T. and Chrispeels, M.J. (1978) Localization of vicilin peptidohydrolase in the cotyledons of mung bean seedling by immuno-fluorescence microscopy. Journal of Cell Biology 79, 10–19.CrossRefGoogle Scholar
Bewley, J.D. and Black, M. (1994) Seeds: Physiology of development and germination, 2nd ed.New York, Plenum Press.CrossRefGoogle Scholar
Chrispeels, M.J., Baumgartner, B. and Harris, N. (1976) Regulation of reserve protein metabolism in the cotyledons of mung bean seedlings. Proceedings of the National Academy of Sciences, USA 73, 3168–3172.CrossRefGoogle ScholarPubMed
DeVries, S., Hoge, H. and Bisseling, T. (1988) Isolation of total and polysomal RNA from plant tissues. pp 1–13in Gelvin, S.B., Schilperoort, R.A. and Verma, D.P.S. (Eds) Plant molecular biology manual. Section B6. Dordrecht, Kluwer Academic Publishers, Section B6.Google Scholar
Ericson, M.C. and Chrispeels, M.J. (1973) Isolation and characterization of glucosamine-containing storage glycoproteins from the cotyledons of Phaseolus aureus. Plant Physiology 52, 98–100.CrossRefGoogle ScholarPubMed
Feinberg, A.P. and Vogelstein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132, 6–13.CrossRefGoogle ScholarPubMed
Hammerton, R.W. and Ho, T.-HD. (1986) Hormonal regulation of the development of protease and carboxypeptidase activities in barley aleurone layers. Plant Physiology 80, 692–697.CrossRefGoogle ScholarPubMed
Herman, E.M., Tague, B.W., Hoffman, L.M., Kjemtrup, S.E. and Chrispeels, M.J. (1990) Retention of phytohemag-glutinin with carboxyterminal tetrapeptide KDEL in the nuclear envelope and the endoplasmic reticulum. Planta 182, 305–312.CrossRefGoogle Scholar
Jones, C.G., Tucker, G.A. and Lycett, G.W. (1996) Pattern of expression and characteristics of a cysteine proteinase cDNA from germinating seeds of pea (Pisum sativum L.). Biochimica et Biophysica Acta 1296, 13–15.CrossRefGoogle ScholarPubMed
Karrer, K.M., Peiffer, S.L. and DiTomas, M.E. (1993) Two distinct gene superfamilies within the family of cysteine protease genes. Proceedings of the National Academy Sciences, USA 90, 3063–3067.CrossRefGoogle Scholar
Kembhavi, A.A., Buttle, D.J., Knight, C.G. and Barrett, A.J. (1993) The two cysteine endopeptidases of legume seeds: purification and characterization by use of specific fluorometric assays. Archives of Biochemistry and Biophysics 303, 208–213.CrossRefGoogle ScholarPubMed
Kern, R. and Chrispeels, M.J. (1978) Influence of the axis on the enzymes of protein and amide metabolism in the cotyledons of mung bean seedlings. Plant Physiology 62, 815–817.CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 277, 680–685.CrossRefGoogle Scholar
Mitsuhashi, W., Koshiba, T. and Minamikawa, T. (1984) Influence of axis removal on amino, carboxy and endopeptidase activities in cotyledons of germinating Vigna mungo seeds. Plant and Cell Physiology 25, 547–554.Google Scholar
Mitsuhashi, W. and Minamikawa, T. (1989) Synthesis and posttranslational activation of sulfhydryl-endopeptidase in cotyledons of germinating Vigna mungo seeds. Plant Physiology 89, 274–279.CrossRefGoogle ScholarPubMed
Muntz, K. (1996) Proteases and proteolytic cleavage of storage proteins in developing and germinating dicotyledonous seeds. Journal of Experimental Botany 47, 605–622.CrossRefGoogle Scholar
Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A laboratory Manual, 2nd Ed.Cold Spring Harbor, NY, Cold Spring Harbor Laboratory Press.Google Scholar
Shutov, A.D. and Vaintraub, I.A. (1987) Degradation of storage proteins in germinating seeds. Phytochemistry 26, 1557–1566.CrossRefGoogle Scholar
Tanaka, T., Minamikawa, T., Yamauchi, D. and Ogushi, Y. (1993) Expression of an endopeptidase (EP-C1) in Phaseolus vulgaris plants. Plant Physiology 101, 421–428.CrossRefGoogle ScholarPubMed
Tanaka, T., Yamauchi, D. and Minamikawa, T. (1991) Nucleotide sequence of cDNA for an endopeptidase (EP-C1) from pods of maturing Phaseolus vulgaris fruits. Plant Molecular Biology 16, 1083–1084.CrossRefGoogle ScholarPubMed
Terasaki, Y., Yamauchi, D., Morikawa, H. and Minamikawa, T. (1995) Influence of axis removal on the expression of a gene for cysteine endopeptidase (EP-C1) in French bean cotyledons during germination. Plant and Cell Physiology 36, 537–541.Google Scholar
Warren, G. (1987) Protein transport: signals and salvage sequences. Nature 327, 17–18.CrossRefGoogle ScholarPubMed
Wilson, K.A. (1986) Role of proteolytic enzymes in the mobilization of protein reserves in the germinating dicot seed. pp 19–47in Dalling, M.J. (Ed.) Plant proteolytic enzymes, Vol. 2. Boca Raton, FL, CRC Press.Google Scholar
Wilson, K.A. and Chen, J.C. (1983) Amino acid sequence of mung bean trypsin inhibitor and its modified forms appearing during germination. Plant Physiology 71, 341–349.CrossRefGoogle ScholarPubMed
Wilson, K.A. and Tan-Wilson, A.L. (1987) Characterization of the proteinase that initiates the degradation of the trypsin inhibitor in germinating mung beans (Vigna radiata). Plant Physiology 84, 93–98.CrossRefGoogle ScholarPubMed
Wilson, K.A., Rightmire, B.R. and Tan-Wilson, A.L. (1985) Involvement of carboxypeptidase in the degradation of the mung bean (Vigna radiata) trypsin inhibitor during germination and early seedling growth. Qualitas Plantarum, Plant Foods in Human Nutrition 35, 195–211.CrossRefGoogle Scholar
Wilson, K.A., Russell, M., Quackenbush, J.F. and Tan-Wilson, A.L. (1995) Characterization of carboxypeptidase I of mung bean seeds. Seed Science Research 5, 209–218.CrossRefGoogle Scholar
Yamaoka, Y., Takeuchi, M. and Morohashi, Y. (1990) Purification and characterization of a cysteine endopeptidase in cotyledons of germinated mung bean seeds. Plant Physiology 94, 561–566.CrossRefGoogle ScholarPubMed
Yamauchi, D., Akasofu, H. and Minamikawa, T. (1992) Cysteine endopeptidase from Vigna mungo: Gene structure and expression. Plant and Cell Physiology 33, 789–797.Google Scholar