Article abstract

Nature Cell Biology 8, 148 - 155 (2006)
Published online: 22 January 2006 | doi:10.1038/ncb1358

Genomic mapping of single-stranded DNA in hydroxyurea-challenged yeasts identifies origins of replication

Wenyi Feng1, David Collingwood2, Max E. Boeck1, Lindsay A. Fox3, Gina M. Alvino1, Walton L. Fangman1, Mosur K. Raghuraman1 & Bonita J. Brewer1

During DNA replication one or both strands transiently become single stranded: first at the sites where initiation of DNA synthesis occurs (known as origins of replication) and subsequently on the lagging strands of replication forks as discontinuous Okazaki fragments are generated. We report a genome-wide analysis of single-stranded DNA (ssDNA) formation in the presence of hydroxyurea during DNA replication in wild-type and checkpoint-deficient rad53 Saccharomyces cerevisiae cells. In wild-type cells, ssDNA was first observed at a subset of replication origins and later 'migrated' bi-directionally, suggesting that ssDNA formation is associated with continuously moving replication forks. In rad53 cells, ssDNA was observed at virtually every known origin, but remained there over time, suggesting that replication forks stall. Telomeric regions seemed to be particularly sensitive to the loss of Rad53 checkpoint function. Replication origins in Schizosaccharomyces pombe were also mapped using our method.

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  1. Department of Genome Sciences, Box 357730, University of Washington, Seattle, WA 98195–7730, USA.
  2. Department of Mathematics, Box 354350, University of Washington, Seattle, WA 98195–7730, USA.
  3. Department of Biology, RC Box 270211, University of Rochester, Rochester, NY 14627–0211, USA.

Correspondence to: Bonita J. Brewer1 e-mail: bbrewer@gs.washington.edu



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