Abstract
Sequence analysis of a 1.33 kb clone from a root cDNA library of Nicotiana rustica revealed an open reading frame encoding a protein of 356 amino acids. The deduced protein has high levels of homology to human cathepsin B protease and a cathepsin B-like cysteine protease from wheat but much lower levels of homology with other plant cysteine proteinases. Southern blotting experiments suggest a limited number of cathepsin B-like genes are present in the genome of N. rustica and also that of N. tabacum. RNA analysis involving a range of tissues, harvested from both Nicotiana species 4–5 h after the beginning of a 16 h photoperiod, revealed the cathepsin B-like gene was being expressed strongly in roots, stem and developing flowers but weakly in mature leaves. Further analysis of RNA extracted from leaf tissue of N. tabacum revealed the gene showed rhythmic expression and also that its expression increased in response to wounding. Analysis of leaf tissues harvested during the latter part of a 16 h photoperiod (11 and 16 h after illumination commenced) showed that transcript levels were two three times higher than in leaf tissue harvested either towards the end of the dark period or 5 h after illumination commenced. When leaf tissue was wounded at 11:00 (5 h after plants were illuminated), and harvested for RNA extraction 6 h later, the level of cathepsin B-like transcript in mesophyll tissue was found to be increased ca. 2-fold relative to the level detected in unwounded controls.
References
Bond JS, Butler PE: Intracellular proteases. Annu Rev Biochem 56: 333–364 (1987).
Carne AC, Moore CH: The amino acid sequence of the tryptic peptides from actinidin, a proleolytic enzyme from the fruit of Actinidia chinensis. Biochem J 173: 73–83 (1978).
Cejudo FJ, Murphy G, Chinoy C, Baulcombe DC: A gibberellin-regulated gene from wheat with sequence homology to cathepsin B of mammalian cells. Plant J 2: 937–948 (1992).
Chan SJ, Segundo BS, McCormick MB, Steiner DF: Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. Proc Natl Acad Sci USA 83: 7721–7725 (1986).
Cohen LW, Coghlan VM, Dihel LC: Sequencing of papain-encoding cDNA. Gene 48: 219–227 (1986).
Goldsbrough PB, Cullis CA: Characterization of the genes for ribosomal RNA in flax. Nucl Acids Res 9: 1301–1309 (1981).
Hamill JD, Parr AJ, Robins RJ, Rhodes MJC: Secondary product formation by cultures of Beta vulgaris and Nicotiana rustica transformed with Agrobacterium rhizogenes. Plant Cell Rep 5: 111–114 (1986).
Kamphuis IG, Drengh J, Baker EN: Thiol proteases. Comparative studies based on the high resolution structures of papain and actinidin and on amino acid sequence information for Cathepsins B and H, and stem Bromelain. J Mol Biol 182: 317–329 (1985).
Linthorst HJM, van der Does C, Brederode FT, Bol JF: Circadian expression and induction by wounding of tobacco genes for cysteine proteinase. Plant Mol Biol 21: 685–694 (1993).
Musil D, Zucic D, Turk D, Engh RA, Mayr I, Huber R, Opovic T, Turk V, Towatari T, Katunuma N, Bode W: The refined 2.15 Å X-ray crystal structure of human liver cathepsin B: the structural basis for its specificity. EMBO J 10: 2321–2330 (1991).
Rogers JC, Dean D, Heck GR: Aleurain: A barley thiol protease closely related to mammalian cathepsin H. Proc Natl Acad Sci USA 82: 6512–6516 (1985).
Vierstra RD: Protein degradation in plants. Annu Rev Plant Physiol Plant Mol Biol 44: 385–410 (1993).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lidgett, A.J., Moran, M., Wong, K.A.L. et al. Isolation and expression pattern of a cDNA encoding a cathepsin B-like protease from Nicotiana rustica . Plant Mol Biol 29, 379–384 (1995). https://doi.org/10.1007/BF00043660
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00043660