@article{7db0a4457a8a445a9fa3158dae93704b,
title = "The human pre-replication complex is an open complex",
abstract = "In eukaryotes, DNA replication initiation requires assembly and activation of the minichromosome maintenance (MCM) 2–7 double hexamer (DH) to melt origin DNA strands. However, the mechanism for this initial melting is unknown. Here, we report a 2.59-{\AA} cryo-electron microscopy structure of the human MCM-DH (hMCM-DH), also known as the pre-replication complex. In this structure, the hMCM-DH with a constricted central channel untwists and stretches the DNA strands such that almost a half turn of the bound duplex DNA is distorted with 1 base pair completely separated, generating an initial open structure (IOS) at the hexamer junction. Disturbing the IOS inhibits DH formation and replication initiation. Mapping of hMCM-DH footprints indicates that IOSs are distributed across the genome in large clusters aligning well with initiation zones designed for stochastic origin firing. This work unravels an intrinsic mechanism that couples DH formation with initial DNA melting to license replication initiation in human cells.",
keywords = "DNA replication initiation, human MCM2–7 complex, initial DNA melting, origin firing, pre-RC, replication licensing",
author = "Jian Li and Jiangqing Dong and Weitao Wang and Daqi Yu and Xinyu Fan and Hui, {Yan Chit} and Lee, {Clare S.K.} and Lam, {Wai Hei} and Nathan Alary and Yang Yang and Yingyi Zhang and Qian Zhao and Chen, {Chun Long} and Tye, {Bik Kwoon} and Shangyu Dang and Yuanliang Zhai",
note = "Funding Information: We thank the Biological Cryo-EM Center at the Hong Kong University of Science and Technology for the data collection of hMCM-DH samples. The center is generously supported by a donation from the Lo Kwee Seong (LKS) Foundation. This work was supported by the Research Grants Council (RGC) of Hong Kong ( GRF16103918 , GRF17112119 , GRF17101720 , GRF17119022 , C7028-19GF , and C7009-20GF to Y. Zhai.; ECS26101919 , GRF16103321 , C7009-20GF , and C6001-21EF to S.D.). S.D. acknowledges support from Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou) ( SMSEGL20SC01-L ), Guangdong Basic and Applied Basic Research Foundation ( 2021A1515012460 ), and Shenzhen Special Fund for Local Science and Technology Development Guided by Central Government ( 2021Szvup140 ). J.D. and D.Y. are supported by LKS fellowships. Work of CLC lab is supported by the YPI program of I. Curie, the ATIP-Avenir program from Centre National de la Recherche Scientifique (CNRS) and Plan Cancer (grant number ATIP/AVENIR : no. 18CT014-00 ); the Agence Nationale de la Recherche (ANR) (grant number ReDeFINe-19-CE12-0016-02 , TELOCHROM-19-CE12-0020-02 ); and Institut National Du Cancer (INCa) (grant number PLBIO19-076 ). W.W. was supported by a COFUND IC-3i International PhD fellowship. Funding Information: We thank the Biological Cryo-EM Center at the Hong Kong University of Science and Technology for the data collection of hMCM-DH samples. The center is generously supported by a donation from the Lo Kwee Seong (LKS) Foundation. This work was supported by the Research Grants Council (RGC) of Hong Kong (GRF16103918, GRF17112119, GRF17101720, GRF17119022, C7028-19GF, and C7009-20GF to Y. Zhai.; ECS26101919, GRF16103321, C7009-20GF, and C6001-21EF to S.D.). S.D. acknowledges support from Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou) (SMSEGL20SC01-L), Guangdong Basic and Applied Basic Research Foundation (2021A1515012460), and Shenzhen Special Fund for Local Science and Technology Development Guided by Central Government (2021Szvup140). J.D. and D.Y. are supported by LKS fellowships. Work of CLC lab is supported by the YPI program of I. Curie, the ATIP-Avenir program from Centre National de la Recherche Scientifique (CNRS) and Plan Cancer (grant number ATIP/AVENIR: no. 18CT014-00); the Agence Nationale de la Recherche (ANR) (grant number ReDeFINe-19-CE12-0016-02, TELOCHROM-19-CE12-0020-02); and Institut National Du Cancer (INCa) (grant number PLBIO19-076). W.W. was supported by a COFUND IC-3i International PhD fellowship. Y. Zhai conceived the study. Y. Zhai, S.D. B.-K.T. and C.-L.C. supervised the project. J.L. X.F. Y.C.H. Y.Y. and Q.Z. constructed cell lines, purified hMCM-DH, and performed functional assays. J.D. S.D. J.L. D.Y. and Y. Zhang. prepared cryo-grids. S.D. J.D. and D.Y. collected data and processed images. J.D. S.D. D.Y. and Y. Zhai built atomic model. J.L. and C.S.K.L. prepared DNA library for deep sequencing. C.-L.C. W.W. N.A. and C.S.K.L. performed bioinformatic analysis, and Y. Zhai, B.-K.T. S.D. C.-L.C. J.D. J.L. D.Y. W.W. and W.H.L. prepared the figures and the manuscript. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",
year = "2023",
month = jan,
day = "5",
doi = "10.1016/j.cell.2022.12.008",
language = "English",
volume = "186",
pages = "98--111.e21",
journal = "Cell",
issn = "0092-8674",
publisher = "Elsevier B.V.",
number = "1",
}