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The replication initiation protein Sld2 regulates helicase assembly.

Authors
  • Bruck, Irina1
  • Kaplan, Daniel L
  • 1 From the Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, Florida 32312.
Type
Published Article
Journal
Journal of Biological Chemistry
Publisher
American Society for Biochemistry and Molecular Biology
Publication Date
Jan 24, 2014
Volume
289
Issue
4
Pages
1948–1959
Identifiers
DOI: 10.1074/jbc.M113.532085
PMID: 24307213
Source
Medline
Keywords
License
Unknown

Abstract

Assembly of the Cdc45-Mcm2-7-GINS (CMG) replicative helicase complex must be regulated to ensure that DNA unwinding is coupled with DNA synthesis. Sld2 is required for the initiation of DNA replication in budding yeast. We identified a mutant of Sld2, Sld2-m1,4, that is specifically defective in Mcm2-7 binding. When this sld2-m1,4 mutant is expressed, cells exhibit severe inhibition of DNA replication. Furthermore, the CMG complex assembles prematurely in G1 in mutant cells, but not wild-type cells. These data suggest that Sld2 binding to Mcm2-7 is essential to block the inappropriate formation of a CMG helicase complex in G1. We also study a mutant of Sld2 that is defective in binding DNA, sld2-DNA, and find that sld2-DNA cells exhibit no GINS-Mcm2-7 interaction. These data suggest that Sld2 association with DNA is required for CMG assembly in S phase.

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