Cooperative binding of GST–dE2F
and GST–dDP to the oligonucleotide AdE2Fwt and competition
by four Draf–E2F oligonucleotides. Radiolabeled double-stranded
AdE2Fwt oligonucleotides were incubated with each lysate from bacteria
carrying pGST-dE2F or pGST-dDP in the absence or presence of the indicated
amounts of competitor oligonucleotides (indicated at the top of
each lane). In the last lane, 2 µl of
lysate from bacteria carrying pGEX-2T alone was introduced. AdE2Fwt,
oligonucleotide containing two wild-type E2F sites from the adenovirus
E2 gene promoter; AdE2Fmut, oligonucleotide containing two mutant
E2F sites from the adenovirus E2 gene promoter; Draf-E2Fsite1,
oligonucleotide containing the E2F site 1 of the D-raf gene
promoter; Draf-E2Fsite2, oligonucleotide containing
the E2F site 2 of the D-raf gene promoter; Draf-E2Fsite3,
oligonucleotide containing the E2F site 3 of the D-raf gene
promoter; Draf-E2Fsite4, oligonucleotide containing
the E2F site 4 of the D-raf gene promoter.