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. 1988 Aug 25;16(16):7961–7973. doi: 10.1093/nar/16.16.7961

The evolutionary conservation of DNA polymerase alpha.

M A Miller 1, D Korn 1, T S Wang 1
PMCID: PMC338503  PMID: 2458565

Abstract

The evolutionary conservation of DNA polymerase alpha was assessed by immunological and molecular genetic approaches. Four anti-human KB cell DNA polymerase alpha monoclonal antibodies were tested for their ability to recognize a phylogenetically broad array of eukaryotic DNA polymerases. While the single non-neutralizing antibody used in this study recognizes higher mammalian (human, simian, canine, and bovine) polymerases only, three neutralizing antibodies exhibit greater, but variable, extents of cross-reactivity among vertebrate species. The most highly cross-reactive antibody recognizes a unique epitope on a 165-180 kDa catalytic polypeptide in cell lysates from several eukaryotic sources, as distant from man as the amphibians. Genomic Southern hybridization studies with the cDNA of the human DNA polymerase alpha catalytic polypeptide identify the existence of many consensus DNA sequences within the DNA polymerase genes of vertebrate, invertebrate, plant and unicellular organisms. These findings illustrate the differential evolutionary conservation of four unique epitopes on DNA polymerase alpha among vertebrates and the conservation of specific genetic sequences, presumably reflective of critical functional domains, in the DNA polymerase genes from a broad diversity of living forms.

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Selected References

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

  1. Chang L. M., Rafter E., Augl C., Bollum F. J. Purification of a DNA polymerase-DNA primase complex from calf thymus glands. J Biol Chem. 1984 Dec 10;259(23):14679–14687. [PubMed] [Google Scholar]
  2. Coen D. M., Furman P. A., Aschman D. P., Schaffer P. A. Mutations in the herpes simplex virus DNA polymerase gene conferring hypersensitivity to aphidicolin. Nucleic Acids Res. 1983 Aug 11;11(15):5287–5297. doi: 10.1093/nar/11.15.5287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  4. Fisher P. A., Korn D. DNA polymerase-alpha. Purification and structural characterization of the near homogeneous enzyme from human KB cells. J Biol Chem. 1977 Sep 25;252(18):6528–6535. [PubMed] [Google Scholar]
  5. Gilead Z., Jeng Y. H., Wold W. S., Sugawara K., Rho H. M., Harter M. L., Green M. Immunological identification of two adenovirus 2-induced early proteins possibly involved in cell transformation. Nature. 1976 Nov 18;264(5583):263–266. doi: 10.1038/264263a0. [DOI] [PubMed] [Google Scholar]
  6. Johnson L. M., Snyder M., Chang L. M., Davis R. W., Campbell J. L. Isolation of the gene encoding yeast DNA polymerase I. Cell. 1985 Nov;43(1):369–377. doi: 10.1016/0092-8674(85)90042-x. [DOI] [PubMed] [Google Scholar]
  7. Kaguni L. S., Rossignol J. M., Conaway R. C., Banks G. R., Lehman I. R. Association of DNA primase with the beta/gamma subunits of DNA polymerase alpha from Drosophila melanogaster embryos. J Biol Chem. 1983 Aug 10;258(15):9037–9039. [PubMed] [Google Scholar]
  8. Kaguni L. S., Rossignol J. M., Conaway R. C., Lehman I. R. Isolation of an intact DNA polymerase-primase from embryos of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2221–2225. doi: 10.1073/pnas.80.8.2221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Larder B. A., Kemp S. D., Darby G. Related functional domains in virus DNA polymerases. EMBO J. 1987 Jan;6(1):169–175. doi: 10.1002/j.1460-2075.1987.tb04735.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Masaki S., Koiwai O., Yoshida S. 10 S DNA polymerase alpha of calf thymus shows a microheterogeneity in its large polypeptide component. J Biol Chem. 1982 Jun 25;257(12):7172–7177. [PubMed] [Google Scholar]
  11. Mechali M., Abadiedebat J., de Recondo A. M. Eukaryotic DNA polymerase alpha. Structural analysis of the enzyme from regenerating rat liver. J Biol Chem. 1980 Mar 10;255(5):2114–2122. [PubMed] [Google Scholar]
  12. Murakami Y., Wobbe C. R., Weissbach L., Dean F. B., Hurwitz J. Role of DNA polymerase alpha and DNA primase in simian virus 40 DNA replication in vitro. Proc Natl Acad Sci U S A. 1986 May;83(9):2869–2873. doi: 10.1073/pnas.83.9.2869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Plevani P., Foiani M., Valsasnini P., Badaracco G., Cheriathundam E., Chang L. M. Polypeptide structure of DNA primase from a yeast DNA polymerase-primase complex. J Biol Chem. 1985 Jun 10;260(11):7102–7107. [PubMed] [Google Scholar]
  14. Quinn J. P., McGeoch D. J. DNA sequence of the region in the genome of herpes simplex virus type 1 containing the genes for DNA polymerase and the major DNA binding protein. Nucleic Acids Res. 1985 Nov 25;13(22):8143–8163. doi: 10.1093/nar/13.22.8143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Tseng B. Y., Ahlem C. N. A DNA primase from mouse cells. Purification and partial characterization. J Biol Chem. 1983 Aug 25;258(16):9845–9849. [PubMed] [Google Scholar]
  17. Tsurumi T., Maeno K., Nishiyama Y. A single-base change within the DNA polymerase locus of herpes simplex virus type 2 can confer resistance to aphidicolin. J Virol. 1987 Feb;61(2):388–394. doi: 10.1128/jvi.61.2.388-394.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wahl A. F., Kowalski S. P., Harwell L. W., Lord E. M., Bambara R. A. Immunoaffinity purification and properties of a high molecular weight calf thymus DNA alpha-polymerase. Biochemistry. 1984 Apr 24;23(9):1895–1899. doi: 10.1021/bi00304a001. [DOI] [PubMed] [Google Scholar]
  19. Wang T. S., Hu S. Z., Korn D. DNA primase from KB cells. Characterization of a primase activity tightly associated with immunoaffinity-purified DNA polymerase-alpha. J Biol Chem. 1984 Feb 10;259(3):1854–1865. [PubMed] [Google Scholar]
  20. Wang T. S., Pearson B. E., Suomalainen H. A., Mohandas T., Shapiro L. J., Schröder J., Korn D. Assignment of the gene for human DNA polymerase alpha to the X chromosome. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5270–5274. doi: 10.1073/pnas.82.16.5270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wong S. W., Wahl A. F., Yuan P. M., Arai N., Pearson B. E., Arai K., Korn D., Hunkapiller M. W., Wang T. S. Human DNA polymerase alpha gene expression is cell proliferation dependent and its primary structure is similar to both prokaryotic and eukaryotic replicative DNA polymerases. EMBO J. 1988 Jan;7(1):37–47. doi: 10.1002/j.1460-2075.1988.tb02781.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yamaguchi M., Tanabe K., Takahashi T., Matsukage A. Chick embryo DNA polymerase alpha. Polypeptide components and their microheterogeneity. J Biol Chem. 1982 Apr 25;257(8):4484–4489. [PubMed] [Google Scholar]

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