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Cdc6 ATPase activity disengages Cdc6 from the pre-replicative complex to promote DNA replication

Journal Article · · eLife
DOI:https://doi.org/10.7554/elife.05795· OSTI ID:1628831
 [1];  [2];  [2];  [3];  [4];  [2];  [5]
  1. Van Andel Research Inst., Grand Rapids, MI (United States); DOE/OSTI
  2. Imperial College London (United Kingdom). Hammersmith Hospital Campus. Faculty of Medicine
  3. Brookhaven National Lab. (BNL), Upton, NY (United States). Biosciences Dept.
  4. Brookhaven National Lab. (BNL), Upton, NY (United States). Biosciences Dept.; Stony Brook Univ., NY (United States). Dept. of Biochemistry and Cell Biology
  5. Van Andel Research Inst., Grand Rapids, MI (United States)
+ Show Author Affiliations
To initiate DNA replication, cells first load an MCM helicase double hexamer at origins in a reaction requiring ORC, Cdc6, and Cdt1, also called pre-replicative complex (pre-RC) assembly. The essential mechanistic role of Cdc6 ATP hydrolysis in this reaction is still incompletely understood. Here, we show that although Cdc6 ATP hydrolysis is essential to initiate DNA replication, it is not essential for MCM loading. Using purified proteins, an ATPase-defective Cdc6 mutant ‘Cdc6-E224Q’ promoted MCM loading on DNA. Cdc6-E224Q also promoted MCM binding at origins in vivo but cells remained blocked in G1-phase. If after loading MCM, Cdc6-E224Q was degraded, cells entered an apparently normal S-phase and replicated DNA, a phenotype seen with two additional Cdc6 ATPase-defective mutants. Cdc6 ATP hydrolysis is therefore required for Cdc6 disengagement from the pre-RC after helicase loading to advance subsequent steps in helicase activation in vivo.
Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
SC0012704
OSTI ID:
1628831
Journal Information:
eLife, Journal Name: eLife Vol. 4; ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.Copyright Statement
Country of Publication:
United States
Language:
English

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Cited By (8)

Ssams2, a Gene Encoding GATA Transcription Factor, Is Required for Appressoria Formation and Chromosome Segregation in Sclerotinia sclerotiorum journal December 2018
Structural basis of Mcm2–7 replicative helicase loading by ORC–Cdc6 and Cdt1 journal February 2017
Deregulated expression of Cdc6 in the skin facilitates papilloma formation and affects the hair growth cycle journal December 2015
MCM interference during licensing of DNA replication in Xenopus egg extracts-Possible Role of a C-terminal region of MCM3- journal January 2018
Yeast heterochromatin regulators Sir2 and Sir3 act directly at euchromatic DNA replication origins journal February 2018
Yeast heterochromatin regulators Sir2 and Sir3 act directly at euchromatic DNA replication origins journal May 2018
Chromosome Duplication in Saccharomyces cerevisiae journal July 2016
MCM interference during licensing of DNA replication in Xenopus egg extracts-Possible Role of a C-terminal region of MCM3- text January 2018

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