FIG. 5.

Ability of E1A 1-80 polypeptide mutants to interfere with complex formation between TBP and TATA DNA. EMSA was performed using as the probe a ³²P-labeled oligonucleotide containing a TATA element. hTBP (TBP) was added (+) as indicated (GST-TBP was used in the lower lanes). E1A 1-80 or mutant E1A 1-80 polypeptide was added as indicated at three concentrations, 50, 100, and 200 ng. The reaction products were analyzed by native polyacrylamide gel electrophoresis and visualized by PhosphorImage analysis as described in Materials and Methods. These analyses were repeated two to three times. Two independent preparations of E1A 1-80 polypeptides exhibiting a mutant phenotype were analyzed with essentially the same results.