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. 1996 Mar;142(3):717–726. doi: 10.1093/genetics/142.3.717

3' -> 5' Exonucleases of DNA Polymerases ε and δ Correct Base Analog Induced DNA Replication Errors on opposite DNA Strands in Saccharomyces Cerevisiae

P V Shcherbakova 1, Y I Pavlov 1
PMCID: PMC1207013  PMID: 8849882

Abstract

The base analog 6-N-hydroxylaminopurine (HAP) induces bidirectional GC -> AT and AT -> GC transitions that are enhanced in DNA polymerase ε and δ 3' -> 5' exonuclease-deficient yeast mutants, pol2-4 and pol3-01, respectively. We have constructed a set of isogenic strains to determine whether the DNA polymerases δ and ε contribute equally to proofreading of replication errors provoked by HAP during leading and lagging strand DNA synthesis. Site-specific GC -> AT and AT -> GC transitions in a Pol(+), pol2-4 or pol3-01 genetic background were scored as reversions of ura3 missense alleles. At each site, reversion was increased in only one proofreading-deficient mutant, either pol2-4 or pol3-01, depending on the DNA strand in which HAP incorporation presumably occurred. Measurement of the HAP-induced reversion frequency of the ura3 alleles placed into chromosome III near to the defined active replication origin ARS306 in two orientations indicated that DNA polymerases ε and δ correct HAP-induced DNA replication errors on opposite DNA strands.

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Selected References

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  1. Abdul-Masih M. T., Bessman M. J. Biochemical studies on the mutagen, 6-N-hydroxylaminopurine. Synthesis of the deoxynucleoside triphosphate and its incorporation into DNA in vitro. J Biol Chem. 1986 Feb 15;261(5):2020–2026. [PubMed] [Google Scholar]
  2. Burgers P. M. Saccharomyces cerevisiae replication factor C. II. Formation and activity of complexes with the proliferating cell nuclear antigen and with DNA polymerases delta and epsilon. J Biol Chem. 1991 Nov 25;266(33):22698–22706. [PubMed] [Google Scholar]
  3. Deshpande A. M., Newlon C. S. The ARS consensus sequence is required for chromosomal origin function in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Oct;12(10):4305–4313. doi: 10.1128/mcb.12.10.4305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Echols H., Goodman M. F. Fidelity mechanisms in DNA replication. Annu Rev Biochem. 1991;60:477–511. doi: 10.1146/annurev.bi.60.070191.002401. [DOI] [PubMed] [Google Scholar]
  5. Gordenin D. A., Malkova A. L., Peterzen A., Kulikov V. N., Pavlov Y. I., Perkins E., Resnick M. A. Transposon Tn5 excision in yeast: influence of DNA polymerases alpha, delta, and epsilon and repair genes. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3785–3789. doi: 10.1073/pnas.89.9.3785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gordenin D. A., Proscyavichus Y. Y., Malkova A. L., Trofimova M. V., Peterzen A. Yeast mutants with increased bacterial transposon Tn5 excision. Yeast. 1991 Jan;7(1):37–50. doi: 10.1002/yea.320070105. [DOI] [PubMed] [Google Scholar]
  7. Kohalmi S. E., Kunz B. A. Role of neighbouring bases and assessment of strand specificity in ethylmethanesulphonate and N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis in the SUP4-o gene of Saccharomyces cerevisiae. J Mol Biol. 1988 Dec 5;204(3):561–568. doi: 10.1016/0022-2836(88)90355-5. [DOI] [PubMed] [Google Scholar]
  8. Kunz B. A., Pierce M. K., Mis J. R., Giroux C. N. DNA sequence analysis of the mutational specificity of u.v. light in the SUP4-o gene of yeast. Mutagenesis. 1987 Nov;2(6):445–453. doi: 10.1093/mutage/2.6.445. [DOI] [PubMed] [Google Scholar]
  9. Lee G. S., Savage E. A., Ritzel R. G., von Borstel R. C. The base-alteration spectrum of spontaneous and ultraviolet radiation-induced forward mutations in the URA3 locus of Saccharomyces cerevisiae. Mol Gen Genet. 1988 Nov;214(3):396–404. doi: 10.1007/BF00330472. [DOI] [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., Bell J. B., Kunkel T. A., Sugino A. Eukaryotic DNA polymerase amino acid sequence required for 3'----5' exonuclease activity. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9473–9477. doi: 10.1073/pnas.88.21.9473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Morrison A., Johnson A. L., Johnston L. H., Sugino A. Pathway correcting DNA replication errors in Saccharomyces cerevisiae. EMBO J. 1993 Apr;12(4):1467–1473. doi: 10.1002/j.1460-2075.1993.tb05790.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Morrison A., Sugino A. The 3'-->5' exonucleases of both DNA polymerases delta and epsilon participate in correcting errors of DNA replication in Saccharomyces cerevisiae. Mol Gen Genet. 1994 Feb;242(3):289–296. doi: 10.1007/BF00280418. [DOI] [PubMed] [Google Scholar]
  14. Newlon C. S., Lipchitz L. R., Collins I., Deshpande A., Devenish R. J., Green R. P., Klein H. L., Palzkill T. G., Ren R. B., Synn S. Analysis of a circular derivative of Saccharomyces cerevisiae chromosome III: a physical map and identification and location of ARS elements. Genetics. 1991 Oct;129(2):343–357. doi: 10.1093/genetics/129.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pavlov Y. I., Noskov V. N., Lange E. K., Moiseeva E. V., Pshenichnov M. R., Khromov-Borisov N. N. The genetic activity of N6-hydroxyadenine and 2-amino-N6-hydroxyadenine in Escherichia coli, Salmonella typhimurium and Saccharomyces cerevisiae. Mutat Res. 1991 Aug;253(1):33–46. doi: 10.1016/0165-1161(91)90343-7. [DOI] [PubMed] [Google Scholar]
  16. Podust V. N., Georgaki A., Strack B., Hübscher U. Calf thymus RF-C as an essential component for DNA polymerase delta and epsilon holoenzymes function. Nucleic Acids Res. 1992 Aug 25;20(16):4159–4165. doi: 10.1093/nar/20.16.4159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Prelich G., Kostura M., Marshak D. R., Mathews M. B., Stillman B. The cell-cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro. Nature. 1987 Apr 2;326(6112):471–475. doi: 10.1038/326471a0. [DOI] [PubMed] [Google Scholar]
  18. Rose M., Winston F. Identification of a Ty insertion within the coding sequence of the S. cerevisiae URA3 gene. Mol Gen Genet. 1984;193(3):557–560. doi: 10.1007/BF00382100. [DOI] [PubMed] [Google Scholar]
  19. Shcherbakova P. V., Pavlov Y. I. Mutagenic specificity of the base analog 6-N-hydroxylaminopurine in the URA3 gene of the yeast Saccharomyces cerevisiae. Mutagenesis. 1993 Sep;8(5):417–421. doi: 10.1093/mutage/8.5.417. [DOI] [PubMed] [Google Scholar]
  20. Simon M., Giot L., Faye G. The 3' to 5' exonuclease activity located in the DNA polymerase delta subunit of Saccharomyces cerevisiae is required for accurate replication. EMBO J. 1991 Aug;10(8):2165–2170. doi: 10.1002/j.1460-2075.1991.tb07751.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sorenson W. G., Simpson J. P., Ong T. M. Comparison of mutagenic and recombinogenic effects of some adenine analogues in Saccharomyces cerevisiae D7. Mutat Res. 1981 Jun;82(1):95–100. doi: 10.1016/0027-5107(81)90141-x. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]

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