Abstract
Centromere-binding protein B (CENP-B) is a widely conserved DNA binding factor associated with heterochromatin and centromeric satellite repeats1. In fission yeast, CENP-B homologues have been shown to silence long terminal repeat (LTR) retrotransposons by recruiting histone deacetylases2. However, CENP-B factors also have unexplained roles in DNA replication3,4. Here we show that a molecular function of CENP-B is to promote replication-fork progression through the LTR. Mutants have increased genomic instability caused by replication-fork blockage that depends on the DNA binding factor switch-activating protein 1 (Sap1), which is directly recruited by the LTR. The loss of Sap1-dependent barrier activity allows the unhindered progression of the replication fork, but results in rearrangements deleterious to the retrotransposon. We conclude that retrotransposons influence replication polarity through recruitment of Sap1 and transposition near replication-fork blocks, whereas CENP-B counteracts this activity and promotes fork stability. Our results may account for the role of LTR in fragile sites, and for the association of CENP-B with pericentromeric heterochromatin and tandem satellite repeats.
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Data deposits
The sequences from the ChIP-seq experiments are deposited in Sequence Read Archive (http://www.ncbi.nlm.nih.gov/sra?term=SRA024710) under accession number SRA024710.
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Acknowledgements
We thank the Martienssen laboratory, V. Aranda, E. Mejia-Ramirez and F. Antequera for technical advice and discussions, and R. Allshire, E. Noguchi, M. O’Connell, P. Espenshade and the National BioResource Project (T. Nakamura, Japan) for strains. This work was supported by National Institutes of Health grant RO1GM076396 to R.A.M., Cancer Research UK grant C9546/A6517 to J.B., l’Agence Nationale de la Recherche grant ANR-06-BLAN-0271 to B.A., a National Health and Medical Research Council C.J. Martin Fellowship to D.V.I. and a Postdoctoral Fellowship from the Spanish Ministry of Education to M.Z.
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M.Z., B.A. and R.A.M. designed the experiments presented and wrote the paper. M.Z. performed and analysed the experiments. M.W.V. provided bioinformatic analysis. D.G. and D.V.I. provided strains. S.W. and J.B. performed additional experiments.
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The file contains Supplementary Table 1, Supplementary Figures 1-11 with legends, Supplementary Methods and additional references. (PDF 9247 kb)
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Zaratiegui, M., Vaughn, M., Irvine, D. et al. CENP-B preserves genome integrity at replication forks paused by retrotransposon LTR. Nature 469, 112–115 (2011). https://doi.org/10.1038/nature09608
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DOI: https://doi.org/10.1038/nature09608
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