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. 1995 Jan 25;23(2):244–247. doi: 10.1093/nar/23.2.244

Structural relationship between DNA polymerases epsilon and epsilon* and their occurrence in eukaryotic cells.

L Uitto 1, J Halleen 1, T Hentunen 1, M Höyhtyä 1, J E Syväoja 1
PMCID: PMC306661  PMID: 7862528

Abstract

Monoclonal antibodies raised against the N-terminal half of human DNA polymerase epsilon bind both to a large > 200 kDa form of DNA polymerase epsilon from HeLa cells and to a small 140 kDa form (DNA polymerase epsilon*) from calf thymus, while antibody against the C-terminal half binds to DNA polymerase epsilon but does not bind to DNA polymerase epsilon*. These results indicate that the two enzymes have common structural motifs in their N-terminal halves, and that DNA polymerase epsilon* is very likely derived from DNA polymerase epsilon by removal of its C-terminal half. DNA polymerase epsilon as well as DNA polymerase epsilon* was detected in extracts from cells of numerous eukaryotic species from yeast to human. The results indicate that DNA polymerase epsilon and its tendency to occur in a smaller form, DNA polymerase epsilon*, are evolutionarily highly conserved and that DNA polymerase epsilon may occur universally in proliferating eukaryotic cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bambara R. A., Jessee C. B. Properties of DNA polymerases delta and epsilon, and their roles in eukaryotic DNA replication. Biochim Biophys Acta. 1991 Jan 17;1088(1):11–24. doi: 10.1016/0167-4781(91)90147-e. [DOI] [PubMed] [Google Scholar]
  2. Crute J. J., Wahl A. F., Bambara R. A. Purification and characterization of two new high molecular weight forms of DNA polymerase delta. Biochemistry. 1986 Jan 14;25(1):26–36. doi: 10.1021/bi00349a005. [DOI] [PubMed] [Google Scholar]
  3. Focher F., Spadari S., Ginelli B., Hottiger M., Gassmann M., Hübscher U. Calf thymus DNA polymerase delta: purification, biochemical and functional properties of the enzyme after its separation from DNA polymerase alpha, a DNA dependent ATPase and proliferating cell nuclear antigen. Nucleic Acids Res. 1988 Jul 25;16(14A):6279–6295. doi: 10.1093/nar/16.14.6279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goscin L. P., Byrnes J. J. DNA polymerase delta: one polypeptide, two activities. Biochemistry. 1982 May 11;21(10):2513–2518. doi: 10.1021/bi00539a034. [DOI] [PubMed] [Google Scholar]
  5. Hamatake R. K., Hasegawa H., Clark A. B., Bebenek K., Kunkel T. A., Sugino A. Purification and characterization of DNA polymerase II from the yeast Saccharomyces cerevisiae. Identification of the catalytic core and a possible holoenzyme form of the enzyme. J Biol Chem. 1990 Mar 5;265(7):4072–4083. [PubMed] [Google Scholar]
  6. Inoue H., Nojima H., Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene. 1990 Nov 30;96(1):23–28. doi: 10.1016/0378-1119(90)90336-p. [DOI] [PubMed] [Google Scholar]
  7. Jessberger R., Podust V., Hübscher U., Berg P. A mammalian protein complex that repairs double-strand breaks and deletions by recombination. J Biol Chem. 1993 Jul 15;268(20):15070–15079. [PubMed] [Google Scholar]
  8. Kesti T., Frantti H., Syväoja J. E. Molecular cloning of the cDNA for the catalytic subunit of human DNA polymerase epsilon. J Biol Chem. 1993 May 15;268(14):10238–10245. [PubMed] [Google Scholar]
  9. Kesti T., Syväoja J. E. Identification and tryptic cleavage of the catalytic core of HeLa and calf thymus DNA polymerase epsilon. J Biol Chem. 1991 Apr 5;266(10):6336–6341. [PubMed] [Google Scholar]
  10. Morrison A., Araki H., Clark A. B., Hamatake R. K., Sugino A. A third essential DNA polymerase in S. cerevisiae. Cell. 1990 Sep 21;62(6):1143–1151. doi: 10.1016/0092-8674(90)90391-q. [DOI] [PubMed] [Google Scholar]
  11. Morrison A., Sugino A. DNA polymerase II, the epsilon polymerase of Saccharomyces cerevisiae. Prog Nucleic Acid Res Mol Biol. 1993;46:93–120. doi: 10.1016/s0079-6603(08)61019-3. [DOI] [PubMed] [Google Scholar]
  12. Nishida C., Reinhard P., Linn S. DNA repair synthesis in human fibroblasts requires DNA polymerase delta. J Biol Chem. 1988 Jan 5;263(1):501–510. [PubMed] [Google Scholar]
  13. Prussak C. E., Almazan M. T., Tseng B. Y. Peptide production from proteins separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis. Anal Biochem. 1989 May 1;178(2):233–238. doi: 10.1016/0003-2697(89)90630-1. [DOI] [PubMed] [Google Scholar]
  14. Siegal G., Turchi J. J., Jessee C. B., Mallaber L. M., Bambara R. A., Myers T. W. Structural relationships between two forms of DNA polymerase epsilon from calf thymus. J Biol Chem. 1992 Feb 25;267(6):3991–3999. [PubMed] [Google Scholar]
  15. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  16. Syvaoja J., Linn S. Characterization of a large form of DNA polymerase delta from HeLa cells that is insensitive to proliferating cell nuclear antigen. J Biol Chem. 1989 Feb 15;264(5):2489–2497. [PubMed] [Google Scholar]
  17. Syväoja J., Suomensaari S., Nishida C., Goldsmith J. S., Chui G. S., Jain S., Linn S. DNA polymerases alpha, delta, and epsilon: three distinct enzymes from HeLa cells. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6664–6668. doi: 10.1073/pnas.87.17.6664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tanaka S., Hu S. Z., Wang T. S., Korn D. Preparation and preliminary characterization of monoclonal antibodies against human DNA polymerase alpha. J Biol Chem. 1982 Jul 25;257(14):8386–8390. [PubMed] [Google Scholar]
  19. Waga S., Stillman B. Anatomy of a DNA replication fork revealed by reconstitution of SV40 DNA replication in vitro. Nature. 1994 May 19;369(6477):207–212. doi: 10.1038/369207a0. [DOI] [PubMed] [Google Scholar]
  20. Wang T. S. Eukaryotic DNA polymerases. Annu Rev Biochem. 1991;60:513–552. doi: 10.1146/annurev.bi.60.070191.002501. [DOI] [PubMed] [Google Scholar]
  21. Wang Z., Wu X., Friedberg E. C. DNA repair synthesis during base excision repair in vitro is catalyzed by DNA polymerase epsilon and is influenced by DNA polymerases alpha and delta in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Feb;13(2):1051–1058. doi: 10.1128/mcb.13.2.1051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weiser T., Gassmann M., Thömmes P., Ferrari E., Hafkemeyer P., Hübscher U. Biochemical and functional comparison of DNA polymerases alpha, delta, and epsilon from calf thymus. J Biol Chem. 1991 Jun 5;266(16):10420–10428. [PubMed] [Google Scholar]

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