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Molecular and Cellular Biology, May 2000, p. 3234-3244, Vol. 20, No. 9
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Cid1, a Fission Yeast Protein Required for S-M
Checkpoint Control when DNA Polymerase
or
Is
Inactivated
Shao-Win
Wang,1
Takashi
Toda,2
Robert
MacCallum,3
Adrian L.
Harris,1 and
Chris
Norbury1,*
Imperial Cancer Research Fund Molecular
Oncology Laboratory, University of Oxford Institute of Molecular
Medicine, John Radcliffe Hospital, Oxford OX3
9DS,1 and Imperial Cancer Research Fund
Cell Regulation Laboratory2 and
Biomolecular Modelling Laboratory,3
London WC2A 3PX, United Kingdom
Received 18 October 1999/Returned for modification 22 December
1999/Accepted 31 January 2000
The S-M checkpoint is an intracellular signaling pathway that
ensures that mitosis is not initiated in cells undergoing DNA replication. We identified cid1, a novel fission yeast
gene, through its ability when overexpressed to confer specific
resistance to a combination of hydroxyurea, which inhibits DNA
replication, and caffeine, which overrides the S-M checkpoint. Cid1
overexpression also partially suppressed the hydroxyurea sensitivity
characteristic of DNA polymerase
mutants and mutants defective in
the "checkpoint Rad" pathway. Cid1 is a member of a family
of putative nucleotidyltransferases including budding yeast Trf4 and
Trf5, and mutation of amino acid residues predicted to be essential for
this activity resulted in loss of Cid1 function in vivo. Two additional
Cid1-like proteins play similar but nonredundant checkpoint-signaling
roles in fission yeast. Cells lacking Cid1 were found to be viable but
specifically sensitive to the combination of hydroxyurea and caffeine
and to be S-M checkpoint defective in the absence of Cds1. Genetic data suggest that Cid1 acts in association with Crb2/Rhp9 and through the
checkpoint-signaling kinase Chk1 to inhibit unscheduled mitosis specifically when DNA polymerase
or
is inhibited.
*
Corresponding author. Mailing address: Imperial Cancer
Research Fund Molecular Oncology Laboratory, University of Oxford
Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3
9DS, United Kingdom. Phone: 44 1865 222415. Fax: 44 1865 222431. E-mail: c.norbury{at}icrf.icnet.uk.
Molecular and Cellular Biology, May 2000, p. 3234-3244, Vol. 20, No. 9
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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