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Mec1p regulates Pds1p levels in S phase: complex coordination of DNA replication and mitosis

Nature Cell Biology volume 3, pages 619–627 (2001)Cite this article

Abstract

Genetic evidence suggests that the securin Pds1p is the target of a late-S-phase checkpoint control. Here we show that Pds1p becomes essential once two-thirds of the genome has been replicated and that the coupling of the completion of genome replication with mitosis relies on the regulation of Pds1p levels. Mec1p is needed to maintain Pds1p levels under S-phase checkpoint conditions. In contrast, Rad53p and Chk1p, needed for the stabilization of Pds1p in the context of the G2 DNA-damage checkpoint pathway, are dispensable. Thus, the Pds1p-dependent late-S-phase checkpoint pathway couples replication with mitosis but is mechanistically distinct from the G2 DNA-damage checkpoint. Finally, we show that the inhibition of spindle elongation in early S phase, controlled by the Mec1p/Rad53p branch, is not regulated via Pds1p/Esp1p. This can mechanistically explain the need for branched S-phase checkpoint controls.

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Figure 1: Pds1p-dependent checkpoint responds to blocked replication forks in late S phase.
Figure 2: Loss of sister-chromatid cohesion and spindle elongation are linked to the extent of ongoing DNA replication in pds1-128 cells.
Figure 3: Pds1p levels persist upon activation of S-phase checkpoint.
Figure 4: Maintenance of Pds1p levels by S-phase checkpoint controls requires Mec1p but not Rad53p or Chk1p.
Figure 5: Spindle elongation in early S phase is independent of Esp1p and Pds1p.

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Acknowledgements

We thank D. Koshland and O. Cohen-Fix for the Pds1p tagging vector, T. Weinert for strains, and P. Russell and N. Rhind for critical reading of the manuscript. D.J.C. was funded by EMBO and US Army Breast Cancer Research fellowships, M.S. by EMBO and HFSP fellowships, and S.J. by the Danish Medical Research Council.

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  1. Sanne Jensen

    Present address: Division of Yeast Genetics, National Institute for Medical Research, Mill Hill, London, NW7 1AA, UK

Authors and Affiliations

  1. Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, 92037, California, USA

    Duncan J. Clarke, Marisa Segal & Steven I. Reed

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  1. Duncan J. Clarke
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  2. Marisa Segal
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Correspondence to Steven I. Reed.

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Clarke, D., Segal, M., Jensen, S. et al. Mec1p regulates Pds1p levels in S phase: complex coordination of DNA replication and mitosis. Nat Cell Biol 3, 619–627 (2001). https://doi.org/10.1038/35083009

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