Dua, Rajiv and Levy, Daniel L. and Campbell, Judith L. (1999) Analysis of the Essential Functions of the C-terminal Protein/Protein Interaction Domain of Saccharomyces cerevisiae pol epsilon and Its Unexpected Ability to Support Growth in the Absence of the DNA Polymerase Domain. Journal of Biological Chemistry, 274 (32). pp. 22283-22288. ISSN 0021-9258. http://resolver.caltech.edu/CaltechAUTHORS:DUAjbc99
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As first observed by Wittenberg (Kesti, T., Flick, K., Keranen, S., Syvaoja, J. E., and Wittenburg, C. (1999) Mol. Cell 3, 679-685), we find that deletion mutants lacking the entire N-terminal DNA polymerase domain of yeast pol epsilon are viable. However, we now show that point mutations in DNA polymerase catalytic residues of pol epsilon are lethal. Taken together, the phenotypes of the deletion and the point mutants suggest that the polymerase of pol epsilon may normally participate in DNA replication but that another polymerase can substitute in its complete absence. Substitution is inefficient because the deletion mutants have serious defects in DNA replication. This observation raises the question of what is the essential function of the C-terminal half of pol epsilon . We show that the ability of the C-terminal half of the polymerase to support growth is disrupted by mutations in the cysteine-rich region, which disrupts both dimerization of the POL2 gene product and interaction with the essential DPB2 subunit, suggesting that this region plays an important architectural role at the replication fork even in the absence of the polymerase function. Finally, the S phase checkpoint, with respect to both induction of RNR3 transcription and cell cycle arrest, is intact in cells where replication is supported only by the C-terminal half of pol epsilon , but it is disrupted in mutants affecting the cysteine-rich region, suggesting that this domain directly affects the checkpoint rather than acting through the N-terminal polymerase active site.
|Additional Information:||Copyright © 1999 by the American Society for Biochemistry and Molecular Biology. (Received for publication, April 22, 1999, and in revised form, June 9, 1999) We are grateful to Curt Wittenberg for communicating his results prior to publication and to Laura Hoopes and Elizabeth Bertani for help with PFGE. This work was supported by Public Health Service Grant 25508 and Grant 1153-F12 from the American Heart Association (to R.D.). The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.|
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