Figure 8.

Vsm1 does not regulate t-SNARE stability. (A) Vsm1 does not affect the stability of Sso. Pulse-chase experiments were performed in sec9-4 cells bearing a control vector (vector), a deletion in VSM1 (vsm1), or overexpressing VSM1 (VSM1). Cells were labeled with [³⁵S]methionine for 3 min and chased with unlabeled methionine/cysteine for up to 180 min. Sso proteins were immunoprecipitated from lysates by using anti-Sso abs and detected in gels by autoradiography. (B) The UBA domain of Vsm1 is not required for the growth suppression of sec9-4 cells. sec9-4 cells overproducing either native Vsm1 (Vsm1) or Vsm1 lacking its UBA domain (ΔUBA) were diluted serially and grown on plates at 26°C and 32°C for 36 h. sec9-4 cells overproducing both Vsm1 and Sso1 (Vsm1/Sso1) were used as a positive control. (C) The UBA domain of Vsm1 is not required for binding to Sso. WT cells overproducing either native Vsm1 (Vsm1) or Vsm1 lacking its UBA domain (ΔUBA) were lysed and subjected to IP by using anti-Vsm1 abs. Proteins were detected using anti-Sso and -Vsm1 abs. TCL (100 μg/lane).