You are here

intact Mcm10 coiled-coil interaction surface is important for origin melting, helicase assembly and the recruitment of Pol-α to Mcm2-7.

Title: An intact Mcm10 coiled-coil interaction surface is important for origin melting, helicase assembly and the recruitment of Pol-α to Mcm2-7.
7 views
0 downloads
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
Date Issued: 2017-05-16
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 α. 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 α 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-α recruitment to the Mcm2-7 complex. We conclude that an intact Mcm10 coiled-coil interaction surface is important for origin melting, helicase assembly, and the recruitment of pol α to Mcm2-7.
Identifier: FSU_pmch_28510759 (IID), 10.1093/nar/gkx438 (DOI), PMC5499591 (PMCID), 28510759 (RID), 28510759 (EID), 3828234 (PII)
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5499591.
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_28510759
Owner Institution: FSU
Is Part Of: Nucleic acids research.
1362-4962

Choose the citation style.
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-α to Mcm2-7. Nucleic Acids Research. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_28510759