FIG. 5.

Dephosphorylation of the Mcm4 subunit prevents the binding of the Mcm complex to pre-Rcs. Chromatin binding assays were performed by incubating sperm nuclei (3,000 sperm heads/μl of extract) in interphase Xenopus extract enriched in the dephosphorylated Mcm4:band-1 isoform or in extract containing equivalent amounts of Mcm4:band-2 and Mcm4:band-1. After 15 min of incubation at room temperature, the chromatin was isolated and the presence of Mcm4 and Cdc6 proteins on the chromatin was detected by Western blot analysis.