Origin licensing requires ATP binding and hydrolysis by the MCM replicative helicase.
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ICR Authors
Authors
Coster, G
Frigola, J
Beuron, F
Morris, EP
Diffley, JFX
Frigola, J
Beuron, F
Morris, EP
Diffley, JFX
Document Type
Journal Article
Date
2014-09-04
Date Accepted
2014-06-26
Abstract
Loading of the six related Minichromosome Maintenance (MCM) proteins as head-to-head double hexamers during DNA replication origin licensing is crucial for ensuring once-per-cell-cycle DNA replication in eukaryotic cells. Assembly of these prereplicative complexes (pre-RCs) requires the Origin Recognition Complex (ORC), Cdc6, and Cdt1. ORC, Cdc6, and MCM are members of the AAA+ family of ATPases, and pre-RC assembly requires ATP hydrolysis. Here we show that ORC and Cdc6 mutants defective in ATP hydrolysis are competent for origin licensing. However, ATP hydrolysis by Cdc6 is required to release nonproductive licensing intermediates. We show that ATP binding stabilizes the wild-type MCM hexamer. Moreover, by analyzing MCM containing mutant subunits, we show that ATP binding and hydrolysis by MCM are required for Cdt1 release and double hexamer formation. This work alters our view of how ATP is used by licensing factors to assemble pre-RCs.
Citation
Molecular cell, 2014, 55 (5), pp. 666 - 677
Source Title
Publisher
CELL PRESS
ISSN
1097-2765
eISSN
1097-4164
Collections
Research Team
Genome Replication
Structural Electron Microscopy
Structural Electron Microscopy