The Ski7 Antiviral Protein Is an EF1-α Homolog That Blocks Expression of Non-Poly(A) mRNA in Saccharomyces cerevisiae

Abstract

ABSTRACT We mapped and cloned SKI7 , a gene that negatively controls the copy number of L-A and M double-stranded RNA viruses in Saccharomyces cerevisiae . We found that it encodes a nonessential 747-residue protein with similarities to two translation factors, Hbs1p and EF1-α. The ski7 mutant was hypersensitive to hygromycin B, a result also suggesting a role in translation. The SKI7 product repressed the expression of nonpolyadenylated [non-poly(A)] mRNAs, whether capped or uncapped, thus explaining why Ski7p inhibits the propagation of the yeast viruses, whose mRNAs lack poly(A). The dependence of the Ski7p effect on 3′ RNA structures motivated a study of the expression of capped non-poly(A) luciferase mRNAs containing 3′ untranslated regions (3′UTRs) differing in length. In a wild-type strain, increasing the length of the 3′UTR increased luciferase expression due to both increased rates and duration of translation. Overexpression of Ski7p efficiently cured the satellite virus M 2 due to a twofold-increased repression of non-poly(A) mRNA expression. Our experiments showed that Ski7p is part of the Ski2p-Ski3p-Ski8p antiviral system because a single ski7 mutation derepresses the expression of non-poly(A) mRNA as much as a quadruple ski2 ski3 ski7 ski8 mutation, and the effect of the overexpression of Ski7p is not obtained unless other SKI genes are functional. ski1/xrn1Δ ski2Δ and ski1/xrn1Δ ski7Δ mutants were viable but temperature sensitive for growth.