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Characterization of a Novel MMS-Sensitive Allele of Schizosaccharomyces pombe mcm4.

ABSTRACT: The minichromosome maintenance (MCM) complex is the conserved helicase motor of the eukaryotic replication fork. Mutations in the Mcm4 subunit are associated with replication stress and double strand breaks in multiple systems. In this work, we characterize a new temperature-sensitive allele of Schizosaccharomyces pombe mcm4⁺ Uniquely among known mcm4 alleles, this mutation causes sensitivity to the alkylation damaging agent methyl methanesulfonate (MMS). Even in the absence of treatment or temperature shift, mcm4-c106 cells show increased repair foci of RPA and Rad52, and require the damage checkpoint for viability, indicating genome stress. The mcm4-c106 mutant is synthetically lethal with mutations disrupting fork protection complex (FPC) proteins Swi1 and Swi3. Surprisingly, we found that the deletion of rif1⁺ suppressed the MMS-sensitive phenotype without affecting temperature sensitivity. Together, these data suggest that mcm4-c106 destabilizes replisome structure.

SUBMITTER: Ranatunga NS

PROVIDER: S-EPMC5068930 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

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Characterization of a Novel MMS-Sensitive Allele of Schizosaccharomyces pombe mcm4.

Ranatunga Nimna S NS Forsburg Susan L SL

G3 (Bethesda, Md.) 20161013 10

The minichromosome maintenance (MCM) complex is the conserved helicase motor of the eukaryotic replication fork. Mutations in the Mcm4 subunit are associated with replication stress and double strand breaks in multiple systems. In this work, we characterize a new temperature-sensitive allele of Schizosaccharomyces pombe mcm4<sup>+</sup> Uniquely among known mcm4 alleles, this mutation causes sensitivity to the alkylation damaging agent methyl methanesulfonate (MMS). Even in the absence of treatm ...[more]

PMID: 27473316

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