Ubiquitination and localization of Ssk1.

<p>(A) Wild-type cells transformed with vector control (pRS415) or with a plasmid expressing Myc-Ssk1 were exponentially grown in HC-Leu (Exp), and subjected to glucose starvation for 30 min (-Glc). Myc-Ssk1 was immunoprecipitated with anti-Myc antibody and ubiquitinated Ssk1 was detected by Western blotting using anti-Ub antibodies. Equal loading of each lane was confirmed by reprobing with anti-Myc antibodies and by immunoblotting with anti-G6pd antibodies. * nonspecific band. (B) Localization of Ssk1-GFP. Wild-type cells expressing Ssk1-GFP were exponentially grown in HC-Leu (EXP), and then starved for glucose for 30 min (-Glc) or subjected to osmotic stress (+NaCl) for 5 min. A strain expressing Hog1-GFP was used as positive control. Intracellular localization of Ssk1-GFP and Hog1-GFP was determined by fluorescence microscopy.