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Molecular and Cellular Biology, August 2009, p. 4025-4032, Vol. 29, No. 15
0270-7306/09/$08.00+0     doi:10.1128/MCB.00185-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cyclin-Dependent Kinase Inhibits Reinitiation of a Normal S-Phase Program during G2 in Fission Yeast{triangledown} ,{dagger}

Lee Kiang,1*,{ddagger} Christian Heichinger,1,{ddagger},§ Stephen Watt,2,¶ Jürg Bähler,2,|| and Paul Nurse1

Laboratory of Yeast Genetics and Cell Biology, The Rockefeller University, New York, New York,1 Cancer Research UK Fission Yeast Functional Genomics Group, Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom2

Received 9 February 2009/ Returned for modification 8 March 2009/ Accepted 22 May 2009

To achieve faithful replication of the genome once in each cell cycle, reinitiation of S phase is prevented in G2 and origins are restricted from refiring within S phase. We have investigated the block to rereplication during G2 in fission yeast. The DNA synthesis that occurs when G2/M cyclin-dependent kinase (CDK) activity is depleted has been assumed to be repeated rounds of S phase without mitosis, but this has not been demonstrated to be the case. We show here that on G2/M CDK depletion in G2, repeated S phases are induced, which are correlated with normal G1/S transcription and attainment of doublings in cell size. Mostly normal mitotic S-phase origins are utilized, although at different efficiencies, and replication is essentially equal across the genome. We conclude that CDK inhibits reinitiation of S phase during G2, and if G2/M CDK is depleted, replication results from induction of a largely normal S-phase program with only small differences in origin usage and efficiency.

* Corresponding author. Mailing address: Laboratory of Yeast Genetics and Cell Biology, The Rockefeller University, 1230 York Avenue, P.O. Box 5, New York, NY 10065. Phone: (212) 327 8475. Fax: (212) 327 8470. E-mail: lkiang{at}rockefeller.edu

{triangledown} Published ahead of print on 1 June 2009.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} These authors contributed equally to this work.

§ Present address: Institute of Plant Biology, Zollikerstrasse 107, 8008 Zürich, Switzerland.

Present address: Regulatory Systems Biology Laboratory, Cancer Research UK Cambridge Research Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom.

|| Present address: Department of Genetics, Evolution and Environment and UCL Cancer Institute, University College London, London WC1E 6BT, United Kingdom.

Molecular and Cellular Biology, August 2009, p. 4025-4032, Vol. 29, No. 15
0270-7306/09/$08.00+0     doi:10.1128/MCB.00185-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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