S. pombe Rtf2 is important for replication fork barrier activity of RTS1 via splicing of Rtf1

Abstract

Arrested replication forks, when restarted by homologous recombination, result in error-prone DNA syntheses and non-allelic homologous recombination. Fission yeast RTS1 is a model fork barrier used to probe mechanisms of recombination-dependent restart. RTS1 barrier activity is entirely dependent on the DNA binding protein Rtf1 and partially dependent on a second protein, Rtf2. Human RTF2 was recently implicated in fork restart, leading us to examine fission yeast Rtf2's role in more detail. In agreement with previous studies, we observe reduced barrier activity upon rtf2 deletion. However, we identified Rtf2 to be physically associated with mRNA processing and splicing factors and rtf2 deletion to cause increased intron retention. One of the most affected introns resided in the rtf1 transcript. Using an intronless rtf1 we observed no reduction in RFB activity in the absence of Rtf2. Thus, Rtf2 is essential for correct rtf1 splicing to allow optimal RTS1 barrier activity.

Data availability

Sequence data is available under GEO dataset GSE192344.

The following data sets were generated

Article and author information

Author details

  1. Alice M Budden

    Genome Damage and Stability Centre, University of Sussex, Brighton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Murat Eravci

    Department of Biochemistry and Biomedicine, University of Sussex, Brighton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Adam T Watson

    Genome Damage and Stability Centre, University of Sussex, Brighton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Eduard Campillo-Funollet

    Genome Damage and Stability Centre, University of Sussex, Brighton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  5. Antony W Oliver

    Genome Damage and Stability Centre, University of Sussex, Brighton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2912-8273
  6. Karel Naiman

    Genome Damage and Stability Centre, University of Sussex, Brighton, United Kingdom
    For correspondence
    karel.naiman@inserm.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3594-1516
  7. Antony M Carr

    Genome Damage and Stability Centre, University of Sussex, Brighton, United Kingdom
    For correspondence
    A.M.Carr@sussex.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2028-2389

Funding

Wellcome Trust (11047/Z/15/Z)

  • Antony M Carr

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Akira Shinohara, Osaka University, Japan

Version history

  1. Preprint posted: March 10, 2022 (view preprint)
  2. Received: March 11, 2022
  3. Accepted: August 19, 2023
  4. Accepted Manuscript published: August 24, 2023 (version 1)
  5. Version of Record published: September 1, 2023 (version 2)

Copyright

© 2023, Budden et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Alice M Budden
  2. Murat Eravci
  3. Adam T Watson
  4. Eduard Campillo-Funollet
  5. Antony W Oliver
  6. Karel Naiman
  7. Antony M Carr
(2023)
S. pombe Rtf2 is important for replication fork barrier activity of RTS1 via splicing of Rtf1
eLife 12:e78554.
https://doi.org/10.7554/eLife.78554

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