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The Escherichia coli Tus–Ter replication fork barrier causes site-specific DNA replication perturbation in yeast

Author

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  • Nicolai B. Larsen

    (Nordea Center for Healthy Aging, Panum Institute 18.1, University of Copenhagen)

  • Ehud Sass

    (Nordea Center for Healthy Aging, Panum Institute 18.1, University of Copenhagen
    Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford
    Present address: Structural Biology Department, Weizmann Institute of Science, Rehovot 7610001, Israel)

  • Catherine Suski

    (Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford
    Present address: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5A, 02 106 Warsaw, Poland)

  • Hocine W. Mankouri

    (Nordea Center for Healthy Aging, Panum Institute 18.1, University of Copenhagen
    Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford)

  • Ian D. Hickson

    (Nordea Center for Healthy Aging, Panum Institute 18.1, University of Copenhagen
    Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford)

Abstract

Replication fork (RF) pausing occurs at both ‘programmed’ sites and non-physiological barriers (for example, DNA adducts). Programmed RF pausing is required for site-specific DNA replication termination in Escherichia coli, and this process requires the binding of the polar terminator protein, Tus, to specific DNA sequences called Ter. Here, we demonstrate that Tus–Ter modules also induce polar RF pausing when engineered into the Saccharomyces cerevisiae genome. This heterologous RF barrier is distinct from a number of previously characterized, protein-mediated, RF pause sites in yeast, as it is neither Tof1-dependent nor counteracted by the Rrm3 helicase. Although the yeast replisome can overcome RF pausing at Tus–Ter modules, this event triggers site-specific homologous recombination that requires the RecQ helicase, Sgs1, for its timely resolution. We propose that Tus–Ter can be utilized as a versatile, site-specific, heterologous DNA replication-perturbing system, with a variety of potential applications.

Suggested Citation

  • Nicolai B. Larsen & Ehud Sass & Catherine Suski & Hocine W. Mankouri & Ian D. Hickson, 2014. "The Escherichia coli Tus–Ter replication fork barrier causes site-specific DNA replication perturbation in yeast," Nature Communications, Nature, vol. 5(1), pages 1-10, May.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms4574
    DOI: 10.1038/ncomms4574
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    Cited by:

    1. Nagham Ghaddar & Yves Corda & Pierre Luciano & Martina Galli & Ylli Doksani & Vincent Géli, 2023. "The COMPASS subunit Spp1 protects nascent DNA at the Tus/Ter replication fork barrier by limiting DNA availability to nucleases," Nature Communications, Nature, vol. 14(1), pages 1-17, December.

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