Structural basis of MCM2-7 replicative helicase loading by ORC-Cdc6 and Cdt1

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Title: Structural basis of MCM2-7 replicative helicase loading by ORC-Cdc6 and Cdt1
Author(s): Yuan, Z
Riera, A
Bai, L
Sun, J
Nandi, S
Spanos, C
Chen, ZA
Barbon, M
Rappsilber, J
Stillman, B
Speck, C
Li, H
Item Type: Journal Article
Abstract: To start DNA replication, the Origin Recognition Complex (ORC) and Cdc6 load a Mcm2-7 double hexamer onto DNA. Without ATP hydrolysis, ORC-Cdc6 recruits one Cdt1-bound Mcm2-7 hexamer, forming an ORC-Cdc6-Cdt1-Mcm2-7 (OCCM) helicase loading intermediate. Here we report a 3.9Å structure of the OCCM on DNA. Flexible Mcm2-7 winged-helix domains (WHD) engage ORC-Cdc6. A three-domain Cdt1 configuration embraces Mcm2, Mcm4, and Mcm6, nearly half of the hexamer. The Cdt1 C-terminal domain extends to the Mcm6 WHD, which binds Orc4 WHD. DNA passes through the ORC-Cdc6 and Mcm2-7 rings. Origin DNA interaction is mediated by an a-helix in Orc4 and positively charged loops in Orc2 and Cdc6. The Mcm2-7 C-tier AAA+ ring is topologically closed by a Mcm5 loop that embraces Mcm2, but the N-tier ring Mcm2-Mcm5 interface remains open. This structure suggests loading mechanics of the first Cdt1-bound Mcm2-7 hexamer by ORC-Cdc6.
Publication Date: 13-Feb-2017
Date of Acceptance: 9-Jan-2017
URI: http://hdl.handle.net/10044/1/43837
DOI: https://dx.doi.org/10.1038/nsmb.3372
ISSN: 1545-9993
Publisher: Nature Publishing Group
Start Page: 316
End Page: 324
Journal / Book Title: Nature Structural & Molecular Biology
Volume: 24
Copyright Statement: © 2017 Nature America, Inc., part of Springer Nature. All rights reserved.
Sponsor/Funder: Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: 107903/Z/15/Z
BB/N000323/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Biophysics
Cell Biology
EUKARYOTIC DNA-REPLICATION
ORIGIN RECOGNITION COMPLEX
CRYO-EM
SACCHAROMYCES-CEREVISIAE
ORC/CDC6/MCM2-7 COMPLEX
QUATERNARY STRUCTURE
PROTEIN COMPLEXES
ATPASE ACTIVITY
BUDDING YEAST
BINDING
Developmental Biology
11 Medical And Health Sciences
03 Chemical Sciences
06 Biological Sciences
Publication Status: Published
Appears in Collections:Clinical Sciences
Molecular Sciences
Faculty of Medicine



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