Purification and functional inactivation of the fission yeast MCMMCM-BP complex

The MCM (mini-chromosome maintenance) complex is the core of the eukaryotic replicative helicase and comprises six proteins, Mcm2-Mcm7. In humans, a variant form of the complex has Mcm2 replaced by the MCM-BP protein. Recent results suggest that a similar complex exists in fission yeast with an essential role in DNA replication and cell cycle progression. Here, we describe the purification and subunit composition of the fission yeast MCMMcb1 complex. Using newly generated temperature-sensitive alleles, we show that loss of MCMMcb1 function leads to accumulation of DNA damage, checkpoint activation and cell cycle arrest, and provide evidence for a role for MCMMcb1 in meiosis. Structured summary of protein interactions: Mcb1 physically interacts with Mcm4 by pull down (View interaction) Mcb1 physically interacts with Mcm5 by pull down (View interaction) Mcb1 physically interacts with Mcm6 by pull down (View interaction) Mcm4 and Mcb1 physically interact by bimolecular fluorescence complementation (View interaction) Mcb1 physically interacts with Mcm3, Mcm4, Mcm5, Mcm6 and Mcm7 by tandem affinity purification (View interaction) Mcb1 physically interacts with Mcm7 by pull down (View interaction) (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.