FIG. 4.

Protein/DNA complexes on ECB elements include Mcm1 and are influenced by sequences flanking the Mcm1 binding site. Shown are gel retardation assays of complexes that form on a 39-bp DNA fragment, including all the sequences around the fourth CLN3 ECB that are protected from DNase I cleavage (lanes 1 to 6) or those that form on the same palindromic site but lack the flanking sequences (lanes 7 and 8). Bar denotes Mcm1-specific complexes. All assays include 0.5 (+) or 1 (++) μl of crude cell extract, except lane 3, which shows the complexes formed with 1 μl of in vitro-translated Mcm1 (ivt). Polyclonal antiserum (Ab) directed against Mcm1 was added (0.5 μl) to lane 2.