Figure 3.

(A) RNA3 stabilization by factor 1a protein in wt and ded1i yeast in the absence of factor 2a protein. Western and Northern blot analyses of wt and ded1i yeast expressing wt RNA3 with or without 1a. Northern blots were hybridized with a positive-strand-specific RNA probe from the coat protein gene. Averages from three or more experiments are shown. (B) Accumulation of 1a and 2a proteins expressed from ADH1 (Left)- or GAL1 (Right)-driven mRNAs and wt RNA3 replication in wt and ded1i yeast. Northern blots were hybridized with a positive-strand-specific RNA probe from the coat protein gene. Percent wt RNA4 values are averages of three or more experiments. (C) Schematic diagram of BMV RNA2, 2aADH mRNA, and 2aGAL mRNA shows BMV RNA2-derived (solid bars) and non-BMV (open bars) 5′ and 3′ NCR sequences. (D) Defective 2a but not 1a expression from wt RNA2 and RNA1, respectively. (Left) 2a protein expression from RNA2 in wt and ded1i yeast. A Northern blot was hybridized with a positive-strand-specific RNA probe from the 2a ORF. (Right) Data are similar to Left, except that wt and ded1i yeast expressed 1a from RNA1. Percent wt 1a or 2a values are averages of three or more experiments.