FIGURE 6.

NBD2* degradation and polyubiquitination are Hsp70- and Hsp40-dependent. Cycloheximide chases, as described in the preceding figure legends, were performed in SSA1 and ssa1-45 (A) and HLJYDJ1 and hlj1Δydj1-151 (B) yeast at 37 °C, and W303a and sse1Δ yeast (D) at 26 °C. Data represent the means ± S.E. (error bars) from either three (SSA1), four (YDJ1HLJ1), or three (W303a) independent experiments, each performed in triplicate (*, p < 0.016 (SSA1); *, p < 0.029 (YDJ1HLJ1); *, p < 0.017 (W303a)). C, SSA1 and ssa1-45 yeast expressing NBD2* were shifted to 37 °C for 12 min prior to lysis, NBD2* was precipitated using anti-HA-agarose beads, and the precipitate was analyzed by immunoblotting with either anti-HA-HRP to detect NBD2* or anti-Myc to detect exogenously expressed Myc-ubiquitin. The arrowhead on the αmyc blot denotes where NBD2* migrates, and molecular weight markers (× 10³) are shown to the left.