Mcm10 self-association is mediated by an N-terminal coiled-coil domain

Minichromosome maintenance protein 10 (Mcm10) is an essential eukaryotic DNA-binding replication factor thought to serve as a scaffold to coordinate enzymatic activities within the replisome. Mcm10 appears to function as an oligomer rather than in its monomeric form (or rather than as a monomer). Ho...

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Bibliographic Details
Published in:PloS one 2013-07, Vol.8 (7), p.e70518-e70518
Main Authors: Du, Wenyue, Josephrajan, Ajeetha, Adhikary, Suraj, Bowles, Timothy, Bielinsky, Anja-Katrin, Eichman, Brandt F
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Analysis
Animals
Baking yeast
Biochemistry
Biology
Biophysics
Cell cycle
Coils
Crystal structure
Crystallography
Cyclin-dependent kinases
Deoxyribonucleic acid
Dimerization
Dimers
DNA
DNA biosynthesis
DNA replication
DNA Replication - physiology
Enzymatic activity
Enzymes
Kinases
Leukemia
Mathematical analysis
Minichromosome Maintenance Proteins - chemistry
Minichromosome Maintenance Proteins - genetics
Minichromosome Maintenance Proteins - metabolism
Models, Molecular
Molecular biology
Mutation
Protein Multimerization
Protein Structure, Tertiary
Proteins
Replication
Saccharomyces cerevisiae
Self-association
Sequence Alignment
Trimers
Ultracentrifugation
Xenopus laevis - genetics
Xenopus laevis - metabolism
Xenopus laevis - physiology
Yeast
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Summary:Minichromosome maintenance protein 10 (Mcm10) is an essential eukaryotic DNA-binding replication factor thought to serve as a scaffold to coordinate enzymatic activities within the replisome. Mcm10 appears to function as an oligomer rather than in its monomeric form (or rather than as a monomer). However, various orthologs have been found to contain 1, 2, 3, 4, or 6 subunits and thus, this issue has remained controversial. Here, we show that self-association of Xenopus laevis Mcm10 is mediated by a conserved coiled-coil (CC) motif within the N-terminal domain (NTD). Crystallographic analysis of the CC at 2.4 Å resolution revealed a three-helix bundle, consistent with the formation of both dimeric and trimeric Mcm10 CCs in solution. Mutation of the side chains at the subunit interface disrupted in vitro dimerization of both the CC and the NTD as monitored by analytical ultracentrifugation. In addition, the same mutations also impeded self-interaction of the full-length protein in vivo, as measured by yeast-two hybrid assays. We conclude that Mcm10 likely forms dimers or trimers to promote its diverse functions during DNA replication.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0070518