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. 1999 Mar 16;96(6):2764–2769. doi: 10.1073/pnas.96.6.2764

Figure 4.

Figure 4

Protein–protein interactions. (A) GST-Pho4 pull-down of normal (WT input) and the S53P form (S53P input) of TFIIB. Equal amounts of either GST alone or GST-Pho4 were incubated with either the normal (WT) or the S53P form of TFIIB. Immobilized samples were washed (500 mM NaCl), eluted, and analyzed by Western blot analysis using polyclonal yeast TFIIB antibody as described in Materials and Methods. (B) GST-Pho4 pull-down of the nIIB input and the S53P form (nS53P input) of TFIIB, and cIIB of normal TFIIB. Equal amounts of GST-Pho4 protein were incubated with the indicated input proteins and analyzed by Western blot as described in A. (C) Intramolecular TFIIB interaction. GST-Pho4, GST-nIIB (WT), and GST-nIIB (S53P) were incubated with cIIB and analyzed as described in A. Western blot analysis (TFIIB antibody) was used to confirm that equal amounts of GST-nIIB (lane 6) and GST-nS53P (lane 7) were used in this experiment.