Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation

• DOI: 10.1016/j.molcel.2014.06.033
• Publication Date: 2014-07-31T15:50:44Z
• Authors: Sukhyun Kang; Megan D. Warner; Stephen P. Bell
• Keywords: DNA Replication; 0301 basic medicine; 03 medical and health sciences; Minichromosome Maintenance Proteins; Models, Genetic; Amino Acid Motifs; Replication Origin; Cell Biology; Molecular Biology

The Mcm2-7 replicative helicase is central to all steps of eukaryotic DNA replication. The hexameric ring of Mcm subunits forms six essential ATPases whose contributions to replication initiation remain unclear. Mcm2-7 complexes containing ATPase-motif mutations showed Mcm2-7 ATP binding and hydrolysis are required for helicase loading. Loading-defective Mcm2-7 mutant complexes were defective in initial Mcm2-7 recruitment or Cdt1 release. Comparison with Cdc6 ATPase mutants showed that Cdc6 ATP hydrolysis is not required for helicase loading but instead drives removal of Mcm2-7 complexes that cannot complete loading. A subset of Mcm2-7 ATPase-site mutants completed helicase loading but could not initiate replication. Individual mutants were defective in distinct events during helicase activation, including maintenance of DNA association, recruitment of the GINS helicase activator, and DNA unwinding. Consistent with its heterohexameric structure, our findings show that the six Mcm2-7 ATPase active sites are specialized for different functions during helicase loading and activation.