Skip to main content
. 2005 Jun 30;24(14):2512–2523. doi: 10.1038/sj.emboj.7600713

Figure 3.

Figure 3

In vitro protein complex formation by Mlo3p, Dss1p, Uap56p and Rae1p. Presence or absence of a protein in a binding reaction is indicated by (+) or (−) on top of the respective lanes. Individual protein bands are identified with arrowheads. Molecular weight markers are shown. A cartoon below each gel depicts hypothetical ternary complexes. (A) GST-Dss1p links Mal-Mlo3p and His-Rae1p. Inputs of Mal-Mlo3p, GST-Dss1p and His-Rae1p are shown (lanes 1–3). Mal-Mlo3p binds GST-Dss1p (lane 4) but not His-Rae1p (lane 5). Mal-Mlo3p pulls down His-Rae1p in the presence of GST-Dss1p (lane 6). (B) His-Uap56p links Mal-Mlo3p to His-Rae1p. Inputs are shown (lanes 1–3). Mal-Mlo3p binds His-Uap56p (lane 4) but not His-Rae1p as in (A) (lane 5), but pulls down His-Rae1p in the presence of his-Uap56p (lane 6). (C) His-Rae1p binds His-Uap56p and GST-Dss1p simultaneously. Inputs including GST are shown (lanes 1–4). GST-Dss1p binds His-Rae1p (lane 5) but not His-Uap56p (lane 6). GST-Dss1p brings down His-Uap56p in the presence of His-Rae1p (lane 7). GST alone fails to bind either His-Rae1p or His-Uap56p (lane 8). (D) His-Mlo3p binds GST-Dss1p and His-Uap56p simultaneously. Lanes 1–3: Input of individual proteins. GST-Dss1p binds His-Mlo3p directly (lane 4) but not His-Uap56 (lane 5). GST-Dss1p forms a complex with His-Uap56p via His-Mlo3p (lane 6).