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Research Article

Replication and segregation of an Escherichia coli chromosome with two replication origins

Xindan Wang, Christian Lesterlin, Rodrigo Reyes-Lamothe, Graeme Ball, and David J. Sherratt
  1. Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom

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PNAS June 28, 2011 108 (26) E243-E250; https://doi.org/10.1073/pnas.1100874108
Xindan Wang
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom
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Christian Lesterlin
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom
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Rodrigo Reyes-Lamothe
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom
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Graeme Ball
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom
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David J. Sherratt
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom
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  • For correspondence: david.sherratt@bioch.ox.ac.uk
  1. Edited by Nancy E. Kleckner, Harvard University, Cambridge, MA, and approved May 17, 2011 (received for review January 18, 2011)

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Abstract

Characterized bacteria, unlike eukaryotes and some archaea, initiate replication bidirectionally from a single replication origin contained within a circular or linear chromosome. We constructed Escherichia coli cells with two WT origins separated by 1 Mb in their 4.64-Mb chromosome. Productive bidirectional replication initiated synchronously at both spatially separate origins. Newly replicated DNA from both origins was segregated sequentially as replication progressed, with two temporally and spatially separate replication termination events. Replication initiation occurred at a cell volume identical to that of cells with a single WT origin, showing that initiation control is independent of cellular and chromosomal oriC concentration. Cells containing just the ectopic origin initiated bidirectional replication at the expected cell mass and at the normal cellular location of that region. In all strains, spatial separation of sister loci adjacent to active origins occurred shortly after their replication, independently of whether replication initiated at the normal origin, the ectopic origin, or both origins.

Footnotes

  • ↵1X.W. and C.L. contributed equally to this work.

  • ↵2Present address: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115.

  • ↵3To whom correspondence should be addressed. E-mail: david.sherratt{at}bioch.ox.ac.uk.
  • Author contributions: X.W., C.L., R.R.-L., and D.J.S. designed research; X.W., C.L., and R.R.-L. performed research; G.B. contributed new reagents/analytic tools; X.W., C.L., and R.R.-L. analyzed data; and X.W., C.L., and D.J.S. wrote the paper.

  • The authors declare no conflict of interest.

  • This article is a PNAS Direct Submission.

  • See Author Summary on page 10383.

  • This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1100874108/-/DCSupplemental.

Freely available online through the PNAS open access option.

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Replication and segregation of an Escherichia coli chromosome with two replication origins
Xindan Wang, Christian Lesterlin, Rodrigo Reyes-Lamothe, Graeme Ball, David J. Sherratt
Proceedings of the National Academy of Sciences Jun 2011, 108 (26) E243-E250; DOI: 10.1073/pnas.1100874108

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Replication and segregation of an Escherichia coli chromosome with two replication origins
Xindan Wang, Christian Lesterlin, Rodrigo Reyes-Lamothe, Graeme Ball, David J. Sherratt
Proceedings of the National Academy of Sciences Jun 2011, 108 (26) E243-E250; DOI: 10.1073/pnas.1100874108
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  • Replication and segregation of an Escherichia coli chromosome with two replication origins
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  • Replication and segregation of an Escherichia coli chromosome with two replication origins
    - Jun 13, 2011
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