RRC ID 36016
Author Perez-Arnaiz P, Bruck I, Kaplan DL.
Title Mcm10 coordinates the timely assembly and activation of the replication fork helicase.
Journal Nucleic Acids Res
Abstract Mcm10 is an essential replication factor that is required for DNA replication in eukaryotes. Two key steps in the initiation of DNA replication are the assembly and activation of Cdc45-Mcm2-7-GINS (CMG) replicative helicase. However, it is not known what coordinates helicase assembly with helicase activation. We show in this manuscript, using purified proteins from budding yeast, that Mcm10 directly interacts with the Mcm2-7 complex and Cdc45. In fact, Mcm10 recruits Cdc45 to Mcm2-7 complex in vitro. To study the role of Mcm10 in more detail in vivo we used an auxin inducible degron in which Mcm10 is degraded upon addition of auxin. We show in this manuscript that Mcm10 is required for the timely recruitment of Cdc45 and GINS recruitment to the Mcm2-7 complex in vivo during early S phase. We also found that Mcm10 stimulates Mcm2 phosphorylation by DDK in vivo and in vitro. These findings indicate that Mcm10 plays a critical role in coupling replicative helicase assembly with helicase activation. Mcm10 is first involved in the recruitment of Cdc45 to the Mcm2-7 complex. After Cdc45-Mcm2-7 complex assembly, Mcm10 promotes origin melting by stimulating DDK phosphorylation of Mcm2, which thereby leads to GINS attachment to Mcm2-7.
Volume 44(1)
Pages 315-29
Published 2016-1-8
DOI 10.1093/nar/gkv1260
PII gkv1260
PMID 26582917
PMC PMC4705653
MeSH Cell Cycle Proteins / metabolism Cell Proliferation DNA Helicases / metabolism* DNA Replication* DNA-Binding Proteins / metabolism Minichromosome Maintenance Proteins / metabolism* Multiprotein Complexes Phosphorylation Protein Binding Replication Origin
IF 11.502
Times Cited 15
Yeast BY25924 BY25925 BY25926