Structural mechanism of helicase loading onto replication origin DNA by ORC-Cdc6.

Proceedings of the National Academy of Sciences of the United States of America
Zuanning YuanChristian Speck

Abstract

DNA replication origins serve as sites of replicative helicase loading. In all eukaryotes, the six-subunit origin recognition complex (Orc1-6; ORC) recognizes the replication origin. During late M-phase of the cell-cycle, Cdc6 binds to ORC and the ORC-Cdc6 complex loads in a multistep reaction and, with the help of Cdt1, the core Mcm2-7 helicase onto DNA. A key intermediate is the ORC-Cdc6-Cdt1-Mcm2-7 (OCCM) complex in which DNA has been already inserted into the central channel of Mcm2-7. Until now, it has been unclear how the origin DNA is guided by ORC-Cdc6 and inserted into the Mcm2-7 hexamer. Here, we truncated the C-terminal winged-helix-domain (WHD) of Mcm6 to slow down the loading reaction, thereby capturing two loading intermediates prior to DNA insertion in budding yeast. In "semi-attached OCCM," the Mcm3 and Mcm7 WHDs latch onto ORC-Cdc6 while the main body of the Mcm2-7 hexamer is not connected. In "pre-insertion OCCM," the main body of Mcm2-7 docks onto ORC-Cdc6, and the origin DNA is bent and positioned adjacent to the open DNA entry gate, poised for insertion, at the Mcm2-Mcm5 interface. We used molecular simulations to reveal the dynamic transition from preloading conformers to the loaded conformers in which the...Continue Reading

References

Feb 1, 1996·Journal of Molecular Graphics·W HumphreyK Schulten
Jan 20, 1999·Proceedings of the National Academy of Sciences of the United States of America·M WeinreichB Stillman
Dec 20, 2000·Proceedings of the National Academy of Sciences of the United States of America·T Seki, J F Diffley
Jul 25, 2003·Proceedings of the National Academy of Sciences of the United States of America·Igor N ChesnokovMichael Botchan
Aug 5, 2003·Genes & Development·Madeleine A Palacios DeBeerCatherine A Fox
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Oct 18, 2005·Nature Structural & Molecular Biology·Christian SpeckBruce Stillman
Jan 3, 2006·Molecular Cell·John C W RandellStephen P Bell
Jul 11, 2006·Nature Structural & Molecular Biology·Megan G ClareyMichael Botchan
Feb 23, 2008·The Journal of Physical Chemistry. B·Albert C PanBenoît Roux
Jul 24, 2008·Proceedings of the National Academy of Sciences of the United States of America·Zhiqiang ChenHuilin Li
Apr 7, 2009·Genome Biology·Bernard P DunckerBrendan J McConkey
Apr 29, 2009·Methods : a Companion to Methods in Enzymology·Leonardo G TrabucoKlaus Schulten
Nov 17, 2009·Proceedings of the National Academy of Sciences of the United States of America·Cecile EvrinChristian Speck
Jan 9, 2010·Acta Crystallographica. Section D, Biological Crystallography·Vincent B ChenDavid C Richardson
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsPeter H Zwart
Apr 13, 2010·Acta Crystallographica. Section D, Biological Crystallography·P EmsleyK Cowtan
Mar 8, 2011·Nature Structural & Molecular Biology·Alessandro CostaJames M Berger
Apr 20, 2011·Proceedings of the National Academy of Sciences of the United States of America·Shixuan LiuYingfang Liu
Dec 6, 2011·Nucleic Acids Research·Changdong LiuGuang Zhu
Jul 11, 2012·Proceedings of the National Academy of Sciences of the United States of America·Artem Y LyubimovJames M Berger
Mar 12, 2013·Nature·Jordi FrigolaJohn F X Diffley
Jul 16, 2013·Nature Structural & Molecular Biology·Jingchuan SunHuilin Li
Sep 26, 2013·Methods in Molecular Biology·Charles C David, Donald J Jacobs
Aug 5, 2014·Molecular Cell·Gideon CosterJohn F X Diffley
Aug 5, 2014·Molecular Cell·Sukhyun KangStephen P Bell
Sep 19, 2014·The Journal of Biological Chemistry·Min WuThomas J Kelly
Oct 17, 2014·Genes & Development·Jingchuan SunHuilin Li
Dec 9, 2014·Journal of Structural Biology·Sjors H W Scheres
Mar 13, 2015·Nature·Franziska BleichertJames M Berger
Jul 30, 2015·Nature·Ningning LiNing Gao
Aug 19, 2015·Journal of Structural Biology·Alexis Rohou, Nikolaus Grigorieff
Jul 9, 2013·Journal of Chemical Theory and Computation·Daniel R Roe, Thomas E Cheatham
Jan 18, 2017·Critical Reviews in Biochemistry and Molecular Biology·Matthew W ParkerJames M Berger
Feb 14, 2017·Nature Structural & Molecular Biology·Simina TicauStephen P Bell

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Citations

Jan 27, 2021·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Isaac Kyei Barffour, Desmond Omane Acheampong
Feb 21, 2021·Clinica Chimica Acta; International Journal of Clinical Chemistry·Tian ZengJuan Zou
Mar 21, 2021·Nature Communications·Marina Guerrero-PuigdevallJordi Frigola
Jun 3, 2021·International Journal of Molecular Sciences·Hui Zhang
Mar 31, 2021·Annual Review of Biochemistry·Ilan AttaliJames M Berger
May 17, 2021·DNA Repair·Yo-Chuen Lin, Supriya G Prasanth
Jun 11, 2021·Molecular Therapy : the Journal of the American Society of Gene Therapy·Yingying JiangJianjun Zhang
Oct 12, 2021·Frontiers in Oncology·Chenhui CaiTongwei Chu

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