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Molecular and Cellular Biology, July 2001, p. 4495-4504, Vol. 21, No. 14
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.14.4495-4504.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Schizosaccharomyces pombe Cells Lacking the Amino-Terminal Catalytic Domains of DNA Polymerase Epsilon Are Viable but Require the DNA Damage Checkpoint Control

Wenyi Feng and Gennaro D'Urso*

Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Miami, Florida 33101-6129

Received 14 February 2001/Returned for modification 14 March 2001/Accepted 30 April 2001

In Schizosaccharomyces pombe, the catalytic subunit of DNA polymerase epsilon (Pol varepsilon ) is encoded by cdc20+ and is essential for chromosomal DNA replication. Here we demonstrate that the N-terminal half of Pol varepsilon  that includes the highly conserved polymerase and exonuclease domains is dispensable for cell viability, similar to observations made with regard to Saccharomyces cerevisiae. However, unlike budding yeast, we find that fission yeast cells lacking the N terminus of Pol varepsilon  (cdc20Delta N-term) are hypersensitive to DNA-damaging agents and have a cell cycle delay. Moreover, the viability of cdc20Delta N-term cells is dependent on expression of rad3+, hus1+, and chk1+, three genes essential for the DNA damage checkpoint control. These data suggest that in the absence of the N terminus of Pol varepsilon , cells accumulate DNA damage that must be repaired prior to mitosis. Our observation that S phase occurs more slowly for cdc20Delta N-term cells suggests that DNA damage might result from defects in DNA synthesis. We hypothesize that the C-terminal half of Pol varepsilon  is required for assembly of the replicative complex at the onset of S phase. This unique and essential function of the C terminus is preserved in the absence of the N-terminal catalytic domains, suggesting that the C terminus can interact with and recruit other DNA polymerases to the site of initiation.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, P.O. Box 016129, University of Miami School of Medicine, Miami, FL 33101-6129. Phone: (305) 243-3105. Fax: (305) 243-3064. E-mail: gdurso{at}miami.edu.

Molecular and Cellular Biology, July 2001, p. 4495-4504, Vol. 21, No. 14
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.14.4495-4504.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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