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Fork Reversal and ssDNA Accumulation at Stalled Replication Forks Owing to Checkpoint Defects

Science, 26 July 2002
Vol. 297, Issue 5581, p. 599-602
DOI: 10.1126/science.1074023

Fork Reversal and ssDNA Accumulation at Stalled Replication Forks Owing to Checkpoint Defects

  1. José M. Sogo1,*,
  2. Massimo Lopes2,*,
  3. Marco Foiani2,
  1. 1 Institute of Cell Biology, ETH Hönggerberg, CH-8093 Zürich, Switzerland.
  2. 2 Istituto F.I.R.C. di Oncologia Molecolare, Via Adamello 16, 20141, Milano, Italy, and Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy.

Abstract

Checkpoint-mediated control of replicating chromosomes is essential for preventing cancer. In yeast, Rad53 kinase protects stalled replication forks from pathological rearrangements. To characterize the mechanisms controlling fork integrity, we analyzed replication intermediates formed in response to replication blocks using electron microscopy. At the forks, wild-type cells accumulate short single-stranded regions, which likely causes checkpoint activation, whereas rad53 mutants exhibit extensive single-stranded gaps and hemi-replicated intermediates, consistent with a lagging-strand synthesis defect. Further, rad53 cells accumulate Holliday junctions through fork reversal. We speculate that, in checkpoint mutants, abnormal replication intermediates begin to form because of uncoordinated replication and are further processed by unscheduled recombination pathways, causing genome instability.

  • * These authors contributed equally to this work.

  • To whom correspondence should be addressed. E-mail: foiani{at}ifom-firc.it

  • Received for publication 26 February 2002.
  • Accepted for publication 28 May 2002.

Citation:

J. M. Sogo, M. Lopes, and M. Foiani, Fork Reversal and ssDNA Accumulation at Stalled Replication Forks Owing to Checkpoint Defects. Science 297, 599-602 (2002).

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