Figure 5.

sse1 mutants are unable to rescue ydj1-151 thermosensitivity. (A) Cultures of ydj1-151 yeast transformed with a galactose-inducible expression vector lacking an insert (1), or containing the gene encoding wild-type Sse1p (3), Sse1-1p (4), Sse1-2p (5), or the Sse1p-CTD (6) were spotted onto SC-ura medium supplemented with galactose and incubated for 4 d at the indicated temperatures. Lane 2 contains ydj1-151 yeast with a multicopy, constitutive YDJ1 expression vector (pAV5). (B) Overnight cultures of each strain were grown in SC-ura supplemented with either glucose (D) or galactose (G) at 26°C, cell extracts were prepared, and immunoblots were analyzed using anti-Sse1p, anti-Ydj1p, or anti-Sec61p antiserum. The arrow indicates the migration of full-length Sse1p, and the asterisk denotes the positions of Sse1p CTD (upper band) and a degradation product (lower band). The numbers correspond to those in A.