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. 1979 May;76(5):2316–2320. doi: 10.1073/pnas.76.5.2316

Functional roles of DNA polymerases beta and gamma.

U Hübscher, C C Kuenzle, S Spadari
PMCID: PMC383591  PMID: 287074

Abstract

The physiological functions of DNA polymerases (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7) beta and gamma were investigated by using neuronal nuclei and synaptosomes isolated from rat brain. UV irradiation of neuronal nuclei from 60-day-old rats resulted in a 7- to 10-fold stimulation of DNA repair synthesis attributable to DNA polymerase beta which, at this developmental stage, is virtually the only DNA polymerase present in the nuclei. No repair synthesis could be elicited by treating the nuclei with N-methyl-N-nitrosourea, but this way probably due to the inability of brain tissues to excise alkylated bases from DNA. The role of DNA polymerase gamma was studied in synaptosomes by using a system mimicking in vivo mitochondrial DNA synthesis. By showing that, under these conditions, DNA replication occurs in mitochondria, and exploiting the fact that DNA polymerase gama is the only DNA polymerase present in mitochondria, evidence was obtained for a role of DNA polymerase gamma in mitochondrial DNA replication. Based on these results and on the wealth of literature on DNA polymerase alpha, we conclude that DNA polymerase alpha is mainly responsible for DNA replication in nuclei, DNA polymerase beta is involved in nuclear DNA repair, and DNA polymerase gamma is the mitochondrial replicating enzyme. However, minor roles for DNA polymerase alpha in DNA repair or for DNA polymerase beta in DNA replication cannot be excluded.

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Selected References

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

  1. BALDWIN R. L., SHOOTER E. M. THE ALKALINE TRANSITION OF BU-CONTAINING DNA AND ITS BEARING ON THE REPLICATION OF DNA. J Mol Biol. 1963 Nov;7:511–526. doi: 10.1016/s0022-2836(63)80098-4. [DOI] [PubMed] [Google Scholar]
  2. Berk A. J., Clayton D. A. Mechanism of mitochondrial DNA replication in mouse L-cells: asynchronous replication of strands, segregation of circular daughter molecules, aspects of topology and turnover of an initiation sequence. J Mol Biol. 1974 Jul 15;86(4):801–824. doi: 10.1016/0022-2836(74)90355-6. [DOI] [PubMed] [Google Scholar]
  3. Bertazzoni U., Scovassi A. I., Brun G. M. Chick-embryo DNA polymerase gamma. Identity of gamma-polymerases purified from nuclei and mitochondria. Eur J Biochem. 1977 Dec 1;81(2):237–248. doi: 10.1111/j.1432-1033.1977.tb11945.x. [DOI] [PubMed] [Google Scholar]
  4. Bertazzoni U., Stefanini M., Noy G. P., Giulotto E., Nuzzo F., Falaschi A., Spadari S. Variations of DNA polymerase-alpha and -beta during prolonged stimulation of human lymphocytes. Proc Natl Acad Sci U S A. 1976 Mar;73(3):785–789. doi: 10.1073/pnas.73.3.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bogenhagen D., Clayton D. A. Mechanism of mitochondrial DNA replication in mouse L-cells: kinetics of synthesis and turnover of the initiation sequence. J Mol Biol. 1978 Feb 15;119(1):49–68. doi: 10.1016/0022-2836(78)90269-3. [DOI] [PubMed] [Google Scholar]
  6. Bolden A., Noy G. P., Weissbach A. DNA polymerase of mitochondria is a gamma-polymerase. J Biol Chem. 1977 May 25;252(10):3351–3356. [PubMed] [Google Scholar]
  7. Ciarrocchi G., Linn S. A cell-free assay measuring repair DNA synthesis in human fibroblasts. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1887–1891. doi: 10.1073/pnas.75.4.1887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clayton D. A., Doda J. N., Friedberg E. C. The absence of a pyrimidine dimer repair mechanism in mammalian mitochondria. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2777–2781. doi: 10.1073/pnas.71.7.2777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edenberg H. J., Anderson S., DePamphilis M. L. Involvement of DNA polymerase alpha in simian virus 40 DNA replication. J Biol Chem. 1978 May 10;253(9):3273–3280. [PubMed] [Google Scholar]
  10. Eichler D. C., Wang T. S., Clayton D. A., Korn D. In vitro replication of mitochondrial DNA. Elongation of the endogenous primer sequence in D loop mitochondrial DNA by human DNA polymerase beta. J Biol Chem. 1977 Nov 10;252(21):7888–7893. [PubMed] [Google Scholar]
  11. Harris C. C., Connor R. J., Jackson F. E., Lieberman M. W. Intranuclear distribution of DNA repair synthesis induced by chemical carcinogens or ultraviolet light in human diploid fibroblasts. Cancer Res. 1974 Dec;34(12):3461–3468. [PubMed] [Google Scholar]
  12. Hershey H. V., Werner D. Evidence for non-deoxynucleotide linkers in Ehrlich ascites tumour cell DNA. Nature. 1976 Jul 8;262(5564):148–150. doi: 10.1038/262148a0. [DOI] [PubMed] [Google Scholar]
  13. Hübscher U., Kuenzle C. C., Spadari S. Identity of DNA polymerase gamma from synaptosomal mitochondria and rat-brain nuclei. Eur J Biochem. 1977 Dec 1;81(2):249–258. doi: 10.1111/j.1432-1033.1977.tb11946.x. [DOI] [PubMed] [Google Scholar]
  14. Hübscher U., Kuenzle C. C., Spadari S. Variation of DNA polymerases-alpha, -beta. and -gamma during perinatal tissue growth and differentiation. Nucleic Acids Res. 1977 Aug;4(8):2917–2929. doi: 10.1093/nar/4.8.2917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ikegami S., Taguchi T., Ohashi M., Oguro M., Nagano H., Mano Y. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha. Nature. 1978 Oct 5;275(5679):458–460. doi: 10.1038/275458a0. [DOI] [PubMed] [Google Scholar]
  16. Kleihues P., Bucheler J. Long-term persistence of O6-methylguanine in rat brain DNA. Nature. 1977 Oct 13;269(5629):625–626. doi: 10.1038/269625a0. [DOI] [PubMed] [Google Scholar]
  17. Kuenzle C. C., Knüsel A., Schümperli D. Isolation of neuronal nuclei from rat brain cortex, rat cerebellum, and pigeon forebrain. Methods Cell Biol. 1977;15:89–96. doi: 10.1016/s0091-679x(08)60210-1. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Otto B., Fanning E. DNA polymerase alpha is associated with replicating SV40 nucleoprotein complexes. Nucleic Acids Res. 1978 May;5(5):1715–1728. doi: 10.1093/nar/5.5.1715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vollenweider H. J., Sogo J. M., Koller T. A routine method for protein-free spreading of double- and single-stranded nucleic acid molecules. Proc Natl Acad Sci U S A. 1975 Jan;72(1):83–87. doi: 10.1073/pnas.72.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Waqar M. A., Evans M. J., Huberman J. A. Effect of 2',3'-dideoxythymidine-5'-triphosphate on HeLa cell in vitro DNA synthesis: evidence that DNA polymerase alpha is the only polymerase required for cellular DNA replication. Nucleic Acids Res. 1978 Jun;5(6):1933–1946. doi: 10.1093/nar/5.6.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weissbach A. Eukaryotic DNA polymerases. Annu Rev Biochem. 1977;46:25–47. doi: 10.1146/annurev.bi.46.070177.000325. [DOI] [PubMed] [Google Scholar]
  23. Whittaker V. P., Michaelson I. A., Kirkland R. J. The separation of synaptic vesicles from nerve-ending particles ('synaptosomes'). Biochem J. 1964 Feb;90(2):293–303. doi: 10.1042/bj0900293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wittek R., Müller H. K., Wyler R. Length heterogeneity in the DNA of vaccinia virus is eliminated on cloning the virus. FEBS Lett. 1978 Jun 1;90(1):41–46. doi: 10.1016/0014-5793(78)80293-2. [DOI] [PubMed] [Google Scholar]
  25. Wolfe L. S., Morgan I. G., Gombos G. Isolation of plasma membranes from rat brain. Biochim Biophys Acta. 1971 Sep 14;241(3):737–751. doi: 10.1016/0005-2736(71)90002-2. [DOI] [PubMed] [Google Scholar]

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