The dynamic nature of the human origin recognition complex revealed through five cryoEM structures

Jaremko, M. J., On, K. F., Thomas, D. R., Stillman, B., Joshua-Tor, L. (August 2020) The dynamic nature of the human origin recognition complex revealed through five cryoEM structures. Elife, 9. e58622. ISSN 2050-084x

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Abstract

Genome replication is initiated from specific origin sites established by dynamic events. The Origin Recognition Complex (ORC) is necessary for orchestrating the initiation process by binding to origin DNA, recruiting CDC6, and assembling the MCM replicative helicase on DNA. Here we report five cryoEM structures of the human ORC (HsORC) that illustrate the native flexibility of the complex. The absence of ORC1 revealed a compact, stable complex of ORC2-5. Introduction of ORC1 opens the complex into several dynamic conformations. Two structures revealed dynamic movements of the ORC1 AAA+ and ORC2 winged-helix domains that likely impact DNA incorporation into the ORC core. Additional twist and pinch motions were observed in an open ORC conformation revealing a hinge at the ORC5·ORC3 interface that may facilitate ORC binding to DNA. Finally, a structure of ORC was determined with endogenous DNA bound in the core revealing important differences between human and yeast origin recognition.

Item Type:	Paper
Subjects:	bioinformatics Investigative techniques and equipment > microscopy > Cryo-electron microscopy bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics Investigative techniques and equipment bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification Investigative techniques and equipment > microscopy bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein structure rendering bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
CSHL Authors:	Stillman, Bruce W. Joshua-Tor, Leemor Jaremko, Matt On, Kin Thomas, Dennis R
Communities:	CSHL Cancer Center Program > Gene Regulation and Inheritance Program CSHL Cancer Center Shared Resources > Mass Spectrometry Service CSHL labs > Joshua-Tor lab CSHL labs > Stillman lab
Highlight:	Stillman, Bruce W.
Depositing User:	Matthew Dunn
Date:	18 August 2020
Date Deposited:	17 Nov 2020 21:32
Last Modified:	30 Jan 2024 21:03
PMCID:	PMC7467728
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/39773

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