Purification and functional inactivation of the fission yeast MCM(MCM-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 MCM(Mcb1) complex. Using newly generated temperature-sensitive alleles, we show that loss of MCM(Mcb1) function leads to accumulation of DNA damage, checkpoint activation and cell cycle arrest, and provide evidence for a role for MCM(Mcb1) in meiosis.
Cancer Research UK London Research Institute, London, UK.
Juan-juan
JJ
Li
Jasmin
J
Schnick
Jacqueline
J
Hayles
Stuart A
SA
MacNeill
D000071877:Checkpoint Kinase 1; D002846:Chromatography, Affinity; D015825:Chromosomes, Fungal / Q000235:genetics; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004271:DNA, Fungal / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D013058:Mass Spectrometry; D008540:Meiosis; D046912:Multiprotein Complexes / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D021122:Protein Subunits / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012568:Schizosaccharomyces / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; D021122:Protein Subunits; D029702:Schizosaccharomyces pombe Proteins; C085412:rad52 protein, S pombe; D011494:Protein Kinases; D000071877:Checkpoint Kinase 1