FIG. 4.

FHV RNA replication in trans in Δydj1 yeast. (A) Growth of wt, Δydj1, and complemented (Δydj1+pYDJ1) yeast cells. Yeast cells transformed with pFA-C/HA and pF1_fs were induced in selective medium with 2% raffinose plus 2% galactose, and cell density was measured by spectrophotometry (OD₆₀₀). Note that Δydj1 and the complemented (Δydj1+pYDJ1) yeast cells have the same growth kinetics. The wt yeast were saturated at an OD₆₀₀ of 4 to 5 in selective medium with 2% raffinose and 2% galactose (data not shown). (B) FHV RNA replication in wt, Δydj1, and complemented Δydj1 yeast cells. Yeast cells transformed with pFA-C/HA and pF1_fs were induced for 24 h, and total RNA and protein were isolated from an equal number of cells and analyzed by Northern blotting and immunoblotting, respectively. Note that total RNA and protein accumulation were increased in the Δydj1 strain in both the presence and absence of complementation. We confirmed the genetic deletion of the YDJ1 locus in Δydj1 yeast by PCR (data not shown), and thus the faint bands in lanes 3 and 4 in the Ydj1p blot represent comigrating cellular proteins that cross-reacted with the Ydj1p antiserum, whereas the reduced signal in lane 1 was due to both underloading (note the PGK blot) and replicate variability.