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© The Rockefeller University Press, 0021-9525/2001/9/913 $5.00
The Journal of Cell Biology, Volume 154, Number 5, September 3, 2001 913-924

Activation of mammalian Chk1 during DNA replication arrest

Carmen Feijoo¹, Clare Hall-Jackson¹, Rong Wu², David Jenkins¹, Jane Leitch¹, David M. Gilbert² and Carl Smythe¹

¹ Division of Cell Signaling, School of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom
² Department of Biochemistry and Molecular Biology, State University of New York, Upstate Medical University, Syracuse, NY 14260

Address correspondence to Carl Smythe, Division of Cell Signaling, School of Life Sciences, University of Dundee, MSI/WTB Complex, Dow St., Dundee DD1 5EH, Scotland, UK. Tel.: 44-1382-345-095. Fax: 44-1382-345-893. E-mail: c.g.w.smythe{at}dundee.ac.uk

Checkpoints maintain order and fidelity in the cell cycle by blocking late-occurring events when earlier events are improperly executed. Here we describe evidence for the participation of Chk1 in an intra-S phase checkpoint in mammalian cells. We show that both Chk1 and Chk2 are phosphorylated and activated in a caffeine-sensitive signaling pathway during S phase, but only in response to replication blocks, not during normal S phase progression. Replication block–induced activation of Chk1 and Chk2 occurs normally in ataxia telangiectasia (AT) cells, which are deficient in the S phase response to ionizing radiation (IR). Resumption of synthesis after removal of replication blocks correlates with the inactivation of Chk1 but not Chk2. Using a selective small molecule inhibitor, cells lacking Chk1 function show a progressive change in the global pattern of replication origin firing in the absence of any DNA replication. Thus, Chk1 is apparently necessary for an intra-S phase checkpoint, ensuring that activation of late replication origins is blocked and arrested replication fork integrity is maintained when DNA synthesis is inhibited.

Key Words: S phase; origins; checkpoint; kinase; cancer

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