Binding of human minichromosome maintenance proteins with histone H3

Abstract

Minichromosome maintenance (MCM) proteins play essential roles in eukaryotic DNA replication, but their biochemical properties remain to be determined. We detected in HeLa cell extracts six proteins, CDC47, CDC46/MCM5, Cdc21, P1/MCM3, Mis5, and BM28/MCM2, by their binding to a specific antibody and by partial sequencing. The human homologs of the MCM2 (BM28), Mis5, Cdc21, and CDC47 proteins were tightly bound to a histone- Sepharose column and purified to near homogeneity, whereas the P1/MCM3 and CDC46/MCM5 proteins passed through. Among the four core histones, the human BM28/MCM2, Mis5, Cdc21, and CDC47 proteins had high affinity for histone H3. Immunoprecipitation with anti-Cdc21 antibody revealed that these four MCM proteins form complexes. These results are consistent with the findings that MCM proteins bind with chromatin in vivo.