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Mutants of Ustilago maydis defective in production of one of two polypeptides of KP6 toxin from the preprotoxin
Authors:Jianshi Tao  Idit Ginzberg  Yigal Koltin and Jeremy A Bruenn
Institution:(1) Department of Biological Sciences, SUNY at Buffalo, 14260 Buffalo, NY, USA;(2) Department of Molecular Microbiology and Biotechnology, George Wise Faculty of Life Sciences, Tel Aviv University, 69978 Ramat Aviv, Israel
Abstract:Double-stranded RNA viruses of Ustilago maydis encode secreted killer toxins to which other cells of the same species and closely related species are sensitive. KP6 toxin consists of two polypeptides, agr and beta, produced from a single precursor preprotoxin. In this work, we cloned complementary DNA for the toxin-encoding segment of two of the KP6 nonkiller mutants NK3 and NK13 that secrete the beta and agr polypeptides, respectively. Both sequence analysis of the cDNA clones and in vitro translation of the toxin-encoding double-stranded RNAs showed that both mutants can produce full-length preprotoxins. Cys51 in agr is converted to Arg in NK3 and Thr25 and Lys42 in beta are changed to Pro and Arg, respectively, in NK13. Although agr and beta are encoded in a single prepropolypeptide, only the beta polypeptide is secreted by NK3 and only the agr polypeptide is secreted by NK 13. This differential expression of peptides from one precursor is a unique phenomenon. Neither of the nonsecreted polypeptides accumulated in the cytosol. The possible effects of these mutations on pre-protoxin folding and their consequences for toxin secretion are discussed.
Keywords:Ustilago maydis killer toxin  Double-stranded RNA virus  Killer toxin mutants  Toxin conformation
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