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A multicystatin is induced by drought‐stress in cowpea ( Vigna unguiculata (L.) Walp.) leaves
Author(s) -
Diop Ndeye Ndack,
Kidrič Marjetka,
Repellin Anne,
Gareil Monique,
d'Arcy-Lameta Agnés,
Pham Thi Anh Thu,
Zuily-Fodil Yasmine
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.10.014
Subject(s) - vigna , cystatin , complementary dna , papain , biology , polyclonal antibodies , heterologous expression , abiotic stress , western blot , botany , biochemistry , recombinant dna , cystatin c , gene , enzyme , genetics , antibody , renal function
Cystatins are protein inhibitors of cystein proteinases belonging to the papain family. In cowpea, cystatin‐like polypeptides and a cDNA have been identified from seeds and metabolic functions have been attributed to them. This paper describes VuC1 , a new cystatin cDNA isolated from cowpea leaves ( Vigna unguiculata (L.) Walp.). Sequence analysis revealed a multicystatin structure with two cystatin‐like domains. The recombinant VUC1 protein (rVUC1) was expressed in an heterologous expression system and purified to apparent homogeneity. It appeared to be an efficient inhibitor of papain activity on a chromogenic substrate. Polyclonal antibodies against rVUC1 were obtained. Involvement of the VuC1 cDNA in the cellular response to various abiotic stresses (progressive drought‐stress, dessication and application of exogenous abscissic acid) was studied, using Northern blot and Western blot analysis, in the leaf tissues of cowpea plants corresponding to two cultivars with different capacity to tolerate drought‐stress. Surprisingly, these abiotic stresses induced accumulation of two VuC1 ‐like messages both translated into VUC1‐like polypeptides. Difference in the transcript accumulation patterns was observed between the two cultivars and related to their respective tolerance level. Presence of multiple cystatin‐like polypeptides and their possible involvement in the control of leaf protein degradation by cysteine proteinases is discussed.