Unreplicated DNA remaining from unperturbed S phases passes through mitosis for resolution in daughter cells

To prevent rereplication of genomic segments, the eukaryotic cell cycle is divided into two nonoverlapping phases. During late mitosis and G1 replication origins are “licensed” by loading MCM2-7 double hexamers and during S phase licensed replication origins activate to initiate bidirectional replic...

Full description

Saved in:
Bibliographic Details
Published inProceedings of the National Academy of Sciences - PNAS Vol. 113; no. 39; pp. E5757 - E5764
Main Authors Moreno, Alberto, Carrington, Jamie T., Albergante, Luca, Mamun, Mohammed Al, Haagensen, Emma J., Komseli, Eirini-Stavroula, Gorgoulis, Vassilis G., Newman, Timothy J., Blow, J. Julian
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 27.09.2016
SeriesPNAS Plus
Subjects
Biological Sciences
Bronchi - cytology
Cell cycle
Cell Cycle Proteins - metabolism
Deoxyribonucleic acid
DNA
DNA - metabolism
Epithelial Cells - metabolism
Eukaryotes
Genetic Loci
Genomes
Genomics
HeLa Cells
Histones - metabolism
Humans
Mitosis
Nuclear Proteins - metabolism
PNAS Plus
Proteins
Replication Origin
RNA Interference
S Phase
Tumor Suppressor p53-Binding Protein 1 - metabolism
UFB
DNA replication
cell cycle
53BP1
MCM
Online AccessGet full text

Cover

More Information
Summary:To prevent rereplication of genomic segments, the eukaryotic cell cycle is divided into two nonoverlapping phases. During late mitosis and G1 replication origins are “licensed” by loading MCM2-7 double hexamers and during S phase licensed replication origins activate to initiate bidirectional replication forks. Replication forks can stall irreversibly, and if two converging forks stall with no intervening licensed origin—a “double fork stall” (DFS)—replication cannot be completed by conventional means. We previously showed how the distribution of replication origins in yeasts promotes complete genome replication even in the presence of irreversible fork stalling. This analysis predicts that DFSs are rare in yeasts but highly likely in large mammalian genomes. Here we show that complementary strand synthesis in early mitosis, ultrafine anaphase bridges, and G1-specific p53-binding protein 1 (53BP1) nuclear bodies provide a mechanism for resolving unreplicated DNA at DFSs in human cells. When origin number was experimentally altered, the number of these structures closely agreed with theoretical predictions of DFSs. The 53BP1 is preferentially bound to larger replicons, where the probability of DFSs is higher. Loss of 53BP1 caused hypersensitivity to licensing inhibition when replication origins were removed. These results provide a striking convergence of experimental and theoretical evidence that unreplicated DNA can pass through mitosis for resolution in the following cell cycle.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Edited by James E. Cleaver, University of California, San Francisco, CA, and approved July 6, 2016 (received for review February 26, 2016)
Author contributions: A.M. designed, performed, and optimized experiments on 53BP1 nuclear bodies, UFBs and immunofluorescence, and ChIP-Seq; J.T.C. performed experiments on 53BP1 nuclear bodies, RPA and γ-H2AX foci, mitotic EdU, and clonogenic assays and performed the flow-cytometry experiments; L.A. performed the ChIP-Seq analysis; L.A. and M.A.M. performed the mathematical and computational analyses; A.M. and E.J.H. developed the 3D FACS protocol; E.-S.K. and V.G.G. made the HBEC cells; A.M. and J.T.C. analyzed data; T.J.N. coordinated the theoretical work; J.J.B. led the project; and A.M., J.T.C., and J.J.B. wrote the paper.
2Present address: Northern Institute for Cancer Research, Medical School, Newcastle University, Newcastle upon Tyne NE2 4HH, United Kingdom.
1A.M. and J.T.C. contributed equally to this work.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1603252113