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. 1981 Nov;148(2):618–623. doi: 10.1128/jb.148.2.618-623.1981

Incision and postincision steps of pyrimidine dimer removal in excision-defective mutants of Saccharomyces cerevisiae.

D R Wilcox, L Prakash
PMCID: PMC216247  PMID: 7028721

Abstract

cdc9, a temperature-sensitive mutant defective in polynucleotide deoxyribonucleic acid (DNA) ligase activity, accumulates low-molecular-weight DNA fragments (as measured by sedimentation of DNA in alkaline sucrose gradients) at the nonpermissive temperature after irradiation with ultraviolet light. This phenotype of cdc9 is a sensitive indicator of successful incision during excision repair of dimers. In strains containing excision-defective mutations in any of nine genes in combination with the cdc9 mutation, the absence of low-molecular-weight DNA at the nonpermissive temperature after ultraviolet treatment suggests that these mutants are incision defective, whereas the presence of low-molecular-weight DNA indicates that the mutants are defective in a step after incision. With rad1, rad2, rad3, rad4, and rad10 mutants, the molecular weight of the DNA remained unchanged after ultraviolet irradiation and incubation at the restrictive temperature, despite the presence of the cdc9 mutation; these mutants are therefore incision defective. Low-molecular-weight DNA was observed in rad14 cdc9 and rad16 cdc9 strains. With the rad16 strain, the accumulation of low-molecular-weight DNA correlated with the amount of excision taking place, whereas in the rad14 mutant strain, no evidence of dimer removal was obtained. Therefore, rad14 is likely to be defective in a step after incision.

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Selected References

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

  1. Clark R. W., Wever G. H., Wiberg J. S. High-molecular-weight DNA and the sedimentation coefficient: a new perspective based on DNA from T7 bacteriophage and two novel forms of T4 bacteriophage. J Virol. 1980 Jan;33(1):438–448. doi: 10.1128/jvi.33.1.438-448.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cox B. S., Parry J. M. The isolation, genetics and survival characteristics of ultraviolet light-sensitive mutants in yeast. Mutat Res. 1968 Jul-Aug;6(1):37–55. doi: 10.1016/0027-5107(68)90101-2. [DOI] [PubMed] [Google Scholar]
  3. Doty P., McGill B. B., Rice S. A. THE PROPERTIES OF SONIC FRAGMENTS OF DEOXYRIBOSE NUCLEIC ACID. Proc Natl Acad Sci U S A. 1958 May;44(5):432–438. doi: 10.1073/pnas.44.5.432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goldring E. S., Grossman L. I., Krupnick D., Cryer D. R., Marmur J. The petite mutation in yeast. Loss of mitochondrial deoxyribonucleic acid during induction of petites with ethidium bromide. J Mol Biol. 1970 Sep 14;52(2):323–335. doi: 10.1016/0022-2836(70)90033-1. [DOI] [PubMed] [Google Scholar]
  5. Haseltine W. A., Gordon L. K., Lindan C. P., Grafstrom R. H., Shaper N. L., Grossman L. Cleavage of pyrimidine dimers in specific DNA sequences by a pyrimidine dimer DNA-glycosylase of M. luteus. Nature. 1980 Jun 26;285(5767):634–641. doi: 10.1038/285634a0. [DOI] [PubMed] [Google Scholar]
  6. Hill W. E., Fangman W. L. Scission of Escherichia coli deoxyribonucleic acid in alkali. Biochemistry. 1973 Apr 24;12(9):1772–1774. doi: 10.1021/bi00733a017. [DOI] [PubMed] [Google Scholar]
  7. Johnston L. H., Nasmyth K. A. Saccharomyces cerevisiae cell cycle mutant cdc9 is defective in DNA ligase. Nature. 1978 Aug 31;274(5674):891–893. doi: 10.1038/274891a0. [DOI] [PubMed] [Google Scholar]
  8. Kutter E. M., Wiberg J. S. Degradation of cytosin-containing bacterial and bacteriophage DNA after infection of Escherichia coli B with bacteriophage T4D wild type and with mutants defective in genes 46, 47 and 56. J Mol Biol. 1968 Dec;38(3):395–411. doi: 10.1016/0022-2836(68)90394-x. [DOI] [PubMed] [Google Scholar]
  9. Lauer G. D., Roberts T. M., Klotz L. C. Determination of the nuclear DNA content of Saccharomyces cerevisiae and implications for the organization of DNA in yeast chromosomes. J Mol Biol. 1977 Aug 25;114(4):507–526. doi: 10.1016/0022-2836(77)90175-9. [DOI] [PubMed] [Google Scholar]
  10. Lawrence C. W., Stewart J. W., Sherman F., Christensen R. Specificity and frequency of ultraviolet-induced reversion of an iso-1-cytochrome c ochre mutant in radiation-sensitive strains of yeast. J Mol Biol. 1974 May 5;85(1):137–162. doi: 10.1016/0022-2836(74)90134-x. [DOI] [PubMed] [Google Scholar]
  11. Mortelmans K., Friedberg E. C., Slor H., Thomas G., Cleaver J. E. Defective thymine dimer excision by cell-free extracts of xeroderma pigmentosum cells. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2757–2761. doi: 10.1073/pnas.73.8.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Prakash L. Defective thymine dimer excision in radiation-sensitive mutants rad10 and rad16 of Saccharomyces cerevisiae. Mol Gen Genet. 1977 Apr 29;152(3):125–128. doi: 10.1007/BF00268808. [DOI] [PubMed] [Google Scholar]
  13. Prakash L., Prakash S. Three additional genes involved in pyrimidine dimer removal in Saccharomyces cerevisiae: RAD7, RAD14 and MMS19. Mol Gen Genet. 1979 Nov;176(3):351–359. doi: 10.1007/BF00333097. [DOI] [PubMed] [Google Scholar]
  14. Prakash L. Repair of pyrimidine dimers in nuclear and mitochondrial DNA of yeast irradiated with low doses of ultraviolet light. J Mol Biol. 1975 Nov 15;98(4):781–795. doi: 10.1016/s0022-2836(75)80010-6. [DOI] [PubMed] [Google Scholar]
  15. Prakash L. Repair of pyrimidine dimers in radiation-sensitive mutants rad3, rad4, rad6 and rad9 of Saccharomyces cerevisiae. Mutat Res. 1977 Oct;45(1):13–20. doi: 10.1016/0027-5107(77)90038-0. [DOI] [PubMed] [Google Scholar]
  16. Prakash S., Prakash L., Burke W., Montelone B. A. Effects of the RAD52 Gene on Recombination in SACCHAROMYCES CEREVISIAE. Genetics. 1980 Jan;94(1):31–50. doi: 10.1093/genetics/94.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Radany E. H., Friedberg E. C. A pyrimidine dimer-DNA glycosylase activity associated with the v gene product of bacterophage T4. Nature. 1980 Jul 10;286(5769):182–185. doi: 10.1038/286182a0. [DOI] [PubMed] [Google Scholar]
  18. Resnick M. A., Setlow J. K. Repair of pyrimidine dimer damage induced in yeast by ultraviolet light. J Bacteriol. 1972 Mar;109(3):979–986. doi: 10.1128/jb.109.3.979-986.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Reynolds R. J. Removal of pyrimidine dimers from Saccharomyces cerevisiae nuclear DNA under nongrowth conditions as detected by a sensitive, enzymatic assay. Mutat Res. 1978 Apr;50(1):43–56. doi: 10.1016/0027-5107(78)90059-3. [DOI] [PubMed] [Google Scholar]
  20. Seeberg E. Reconstitution of an Escherichia coli repair endonuclease activity from the separated uvrA+ and uvrB+/uvrC+ gene products. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2569–2573. doi: 10.1073/pnas.75.6.2569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Snow R. Mutants of yeast sensitive to ultraviolet light. J Bacteriol. 1967 Sep;94(3):571–575. doi: 10.1128/jb.94.3.571-575.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Unrau P., Wheatcroft R., Cox B. S. The excision of pyrimidine dimers from DNA of ultraviolet irradiated yeast. Mol Gen Genet. 1971;113(4):359–362. doi: 10.1007/BF00272336. [DOI] [PubMed] [Google Scholar]
  23. Unrau P., Wheatcroft R., Cox B., Olive T. The formation of pyrimidine dimers in the DNA of fungi and bacteria. Biochim Biophys Acta. 1973 Jul 27;312(4):626–632. doi: 10.1016/0005-2787(73)90065-8. [DOI] [PubMed] [Google Scholar]
  24. Waters R., Moustacchi E. The disappearance of ultraviolet-induced pyrimidine dimers from the nuclear DNA of exponential and stationary phase cells of Saccharomyces cerevisiae following various post-irradiation treatments. Biochim Biophys Acta. 1974 Jul 24;353(4):407–419. doi: 10.1016/0005-2787(74)90048-3. [DOI] [PubMed] [Google Scholar]
  25. Youngs D. A., Van der Schueren E., Smith K. C. Separate branches of the uvr gene-dependent excision repair process in ultraviolet-irradiated Escherichia coli K-12 cells; their dependence upon growth medium and the polA, recA, recB, and exrA genes. J Bacteriol. 1974 Feb;117(2):717–725. doi: 10.1128/jb.117.2.717-725.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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