Mutants of Ustilago maydis defective in production of one of two polypeptides of KP6 toxin from the preprotoxin.
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, alpha 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 alpha 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 alpha is converted to Arg in NK3 and Thr25 and Lys42 in beta are changed to Pro and Arg, respectively, in NK13. Although alpha and beta are encoded in a single prepropolypeptide, only the beta polypeptide is secreted by NK3 and only the alpha polypeptide is secreted by NK13. 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 preprotoxin folding and their consequences for toxin secretion are discussed.
Tao
J
J
Department of Biological Sciences, SUNY, Buffalo 14260.
Ginzberg
I
I
Koltin
Y
Y
Bruenn
J A
JA
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009183:Mycotoxins / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D014588:Ustilago / Q000235:genetics*; D014764:Viral Proteins
D004271:DNA, Fungal; D009183:Mycotoxins; D011498:Protein Precursors; C051654:Ustilago maydis toxin p6, Ustilago maydis virus; D014764:Viral Proteins