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Molecular and Cellular Biology, April 2003, p. 2733-2748, Vol. 23, No. 8
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.8.2733-2748.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Saccharomyces cerevisiae DNA Polymerase {varepsilon} and Polymerase {sigma} Interact Physically and Functionally, Suggesting a Role for Polymerase {varepsilon} in Sister Chromatid Cohesion

Shaune Edwards, Caroline M. Li, Daniel L. Levy, Jessica Brown,{dagger} Peter M. Snow, and Judith L. Campbell*

Braun Laboratories, California Institute of Technology, Pasadena, California 91125

Received 16 August 2002/ Returned for modification 25 September 2002/ Accepted 16 January 2003

The large subunit of Saccharomyces cerevisiae DNA polymerase {varepsilon}, Pol2, comprises two essential functions. The N terminus has essential DNA polymerase activity. The C terminus is also essential, but its function is unknown. We report here that the C-terminal domain of Pol2 interacts with polymerase {sigma} (Pol {sigma}), a recently identified, essential nuclear nucleotidyl transferase encoded by two redundant genes, TRF4 and TRF5. This interaction is functional, since Pol {sigma} stimulates the polymerase activity of the Pol {varepsilon} holoenzyme significantly. Since Trf4 is required for sister chromatid cohesion as well as for completion of S phase and repair, the interaction suggested that Pol {varepsilon}, like Pol {sigma}, might form a link between the replication apparatus and sister chromatid cohesion and/or repair machinery. We present evidence that pol2 mutants are defective in sister chromatid cohesion. In addition, Pol2 interacts with SMC1, a subunit of the cohesin complex, and with ECO1/CTF7, required for establishing sister chromatid cohesion; and pol2 mutations act synergistically with smc1 and scc1. We also show that trf5{Delta} mutants, like trf4{Delta} mutants, are defective in DNA repair and sister chromatid cohesion.

* Corresponding author. Mailing address: Braun Laboratories 147-75, California Institute of Technology, Pasadena, CA 91125. Phone: (626) 395-6053. Fax: (626) 405-9452. E-mail: jcampbel{at}its.caltech.edu.

{dagger} Present address: Department of Biochemistry and Biophysics, University of San Francisco, San Francisco, CA 94143.

Molecular and Cellular Biology, April 2003, p. 2733-2748, Vol. 23, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.8.2733-2748.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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