Figure 5.
(A) Effect of Sec10Δ and Sec10CT on the composition of the exocyst complex. Yeast containing c-myc3–tagged Sec8p and expressing Sec10ΔC (lane 2) or Sec10CT (lane 3) after 10 h growth in galactose were labeled with [35S]-cysteine and -methionine. The exocyst complex was precipitated using the c-myc3 tag present on Sec8p as described in MATERIALS AND METHODS. Individual proteins in the exocyst were identified according to an immunoprecipitate from cells containing only c-myc3–tagged Sec8p. The position of Sec10p was further identified by immunoprecipitation from yeast overexpressing Sec10p (lane 1). (B) Sec10ΔC displaces Sec10p from Sec15p. c-myc–tagged Sec10p and Sec15p were translated either separately (left panel) or were cotranslated (right panel) in vitro. After addition of either GST, GST-Sec10ΔC (200 μg/ml final concentration), or no addition, the c-myc–tagged Sec10p was immunoprecipitated with anti–c-myc antibodies.