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Regulation of heterochromatic DNA replication by histone H3 lysine 27 methyltransferases

Nature volume 466, pages 987–991 (2010)Cite this article

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Abstract

Multiple pathways prevent DNA replication from occurring more than once per cell cycle1. These pathways block re-replication by strictly controlling the activity of pre-replication complexes, which assemble at specific sites in the genome called origins. Here we show that mutations in the homologous histone 3 lysine 27 (H3K27) monomethyltransferases, ARABIDOPSIS TRITHORAX-RELATED PROTEIN5 (ATXR5) and ATXR6, lead to re-replication of specific genomic locations. Most of these locations correspond to transposons and other repetitive and silent elements of the Arabidopsis genome. These sites also correspond to high levels of H3K27 monomethylation, and mutation of the catalytic SET domain is sufficient to cause the re-replication defect. Mutation of ATXR5 and ATXR6 also causes upregulation of transposon expression and has pleiotropic effects on plant development. These results uncover a novel pathway that prevents over-replication of heterochromatin in Arabidopsis.

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Figure 1: Heterochromatic DNA is over-produced in atxr5 atxr6 mutants.
Figure 2: Increased heterochromatic DNA in atxr5 atxr6 mutants is consistent with re-replication of chromatin.
Figure 3: Genome-wide mapping of H3K27me1 and anticorrelation with H3K4 methylation.
Figure 4: Functional PHD and SET domains and the PIP motif are required for the regulation of DNA replication by ATXR6.

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Gene Expression Omnibus

Data deposits

Sequencing files have been deposited at GEO (accession codes GSE22411 and GSE21673).

Change history

  • 19 August 2010

    Author Luting Zhuo's family name was corrected for the print issue.

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Acknowledgements

We thank G. Lambert and D. Galbraith for assistance with flow cytometry; Y. Bernatavichute for assistance with ChIP experiments; and M. Pellegrini and S. Cokus for advice on data analyses. Y.J. was supported by a fellowship from Le Fonds Québécois de la Recherche sur la Nature et les Technologies (FQRNT). S.F. is a Howard Hughes Medical Institute Fellow of the Life Science Research Foundation. Research in the Michaels’ laboratory was supported by grants from the National Institutes of Health (GM075060), the Indiana METACyt Initiative of Indiana University, and the Lilly Endowment, Inc. C.G. was supported by grants from the Spanish Ministry of Science and Innovation (BFU2009-9783 and CSD2007-57B). S.E.J. is an investigator of the Howard Hughes Medical Institute.

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Author notes

  1. Yannick Jacob and Hume Stroud: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biology, Indiana University, 915 East Third Street, Bloomington, 47405, Indiana, USA

    Yannick Jacob, Chantal LeBlanc, Luting Zhuo, Christiane Hassel & Scott D. Michaels

  2. Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, Los Angeles, 90095, California, USA

    Hume Stroud, Elena Caro & Steven E. Jacobsen

  3. Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, 90095, California, USA

    Suhua Feng & Steven E. Jacobsen

  4. Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas, Universidad Autonoma de Madrid, Nicolas Cabrera 1, Cantoblanco, Madrid 28049, Spain,

    Crisanto Gutierrez

Authors
  1. Yannick Jacob
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Contributions

S.D.M., S.E.J. and C.G. directed the research. Y.J., H.S., C.L., S.F., L.Z., E.C. and C.H. performed experiments. H.S. analysed data. H.S., Y.J., S.E.J. and S.D.M. prepared the manuscript.

Corresponding authors

Correspondence to Scott D. Michaels or Steven E. Jacobsen.

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The authors declare no competing financial interests.

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This file contains Supplementary Figures 1-7 with legends and Supplementary Tables 1-2. (PDF 2244 kb)

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Jacob, Y., Stroud, H., LeBlanc, C. et al. Regulation of heterochromatic DNA replication by histone H3 lysine 27 methyltransferases. Nature 466, 987–991 (2010). https://doi.org/10.1038/nature09290

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