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Intact Mcm10 Coiled-coil Interaction Surface Is Important For Origin Melting, Helicase Assembly And The Recruitment Of Pol-alpha To Mcm2-7

Title: An Intact Mcm10 Coiled-coil Interaction Surface Is Important For Origin Melting, Helicase Assembly And The Recruitment Of Pol-alpha To Mcm2-7.
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Name(s): Perez-Arnaiz, Patricia, author
Bruck, Irina, author
Colbert, Max K., author
Kaplan, Daniel L., author
Type of Resource: text
Genre: Journal Article
Text
Journal Article
Date Issued: 2017-07-07
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Mcm10 is an essential eukaryotic factor required for DNA replication. The replication fork helicase is composed of Cdc45, Mcm2-7 and GINS (CMG). DDK is an S-phase-specific kinase required for replication initiation, and the DNA primase-polymerase in eukaryotes is pol alpha. Mcm10 forms oligomers in vitro, mediated by the coiled-coil domain at the N-terminal region of the protein. We characterized an Mcm10 mutant at the N-terminal Domain (NTD), Mcm10-4A, defective for self-interaction. We found that the Mcm10-4A mutant was defective for stimulating DDK phosphorylation of Mcm2, binding to eighty-nucleotide ssDNA, and recruiting pol alpha to Mcm2-7 in vitro. Expression of wild-type levels of mcm10-4A resulted in severe growth and DNA replication defects in budding yeast cells, with diminished DDK phosphorylation of Mcm2. We then expressed the mcm10-4A in mcm5-bob1 mutant cells to bypass the defects mediated by diminished stimulation of DDK phosphorylation of Mcm2. Expression of wild-type levels of mcm10-4A in mcm5-bob1 mutant cells resulted in severe growth and DNA replication defects, along with diminished RPA signal at replication origins. We also detected diminished GINS and pol-alpha recruitment to the Mcm2-7 complex. We conclude that an intact Mcm10 coiled-coil interaction surface is important for originmelting, helicase assembly, and the recruitment of pol alpha to Mcm2-7.
Identifier: FSU_libsubv1_wos_000404879000030 (IID), 10.1093/nar/gkx438 (DOI)
Keywords: saccharomyces-cerevisiae, budding yeast, s-phase, single-stranded-dna, structural basis, binding properties, cyclin-dependent kinases, dna-polymerase-alpha, double-hexameric mcm2-7, replication fork helicase
Publication Note: The publisher's version of record is available at https://doi.org/10.1093/nar/gkx438
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_libsubv1_wos_000404879000030
Owner Institution: FSU
Is Part Of: Nucleic Acids Research.
0305-1048
Issue: iss. 12, vol. 45

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Perez-Arnaiz, P., Bruck, I., Colbert, M. K., & Kaplan, D. L. (2017). An Intact Mcm10 Coiled-coil Interaction Surface Is Important For Origin Melting, Helicase Assembly And The Recruitment Of Pol-alpha To Mcm2-7. Nucleic Acids Research. Retrieved from http://purl.flvc.org/fsu/fd/FSU_libsubv1_wos_000404879000030