Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Mar 1;91(5):1642–1646. doi: 10.1073/pnas.91.5.1642

Efficient removal of uracil from G.U mispairs by the mismatch-specific thymine DNA glycosylase from HeLa cells.

P Neddermann 1, J Jiricny 1
PMCID: PMC43219  PMID: 8127859

Abstract

The uracil DNA glycosylases (EC 3.2.2.3) characterized to date remove uracil from DNA irrespective of whether it is base paired with adenine or mispaired with guanine in double-stranded substrates or whether it is found in single-stranded DNA. We report here the characterization of uracil glycosylase activity that can remove the base solely from a mispair with guanine. It does not recognize uracil either in A.U pairs or in single-stranded substrates. The enzyme, a 55-kDa polypeptide, was previously characterized as a mismatch-specific thymine DNA glycosylase and was thought to be responsible solely for the correction (to G.C) of G.T mispairs that arise as a result of spontaneous hydrolytic deamination of 5-methylcytosine to thymine. Given the broader substrate specificity of the enzyme (in addition to uracil and thymine, the protein can also remove 5-bromouracil from mispairs with guanine), we propose that its biological role in vivo may also include the correction of a subset of G.U mispairs inefficiently removed by the more abundant ubiquitous uracil glycosylases, such as those arising from cytosine deamination in G+C-rich regions of the genome.

Full text

PDF
1642

Images in this article

1643Image
on p.1643
1644Image
on p.1644
1645Image
on p.1645
1645Image
on p.1645

Selected References

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

  1. Bennett S. E., Mosbaugh D. W. Characterization of the Escherichia coli uracil-DNA glycosylase.inhibitor protein complex. J Biol Chem. 1992 Nov 5;267(31):22512–22521. [PubMed] [Google Scholar]
  2. Boiteux S., Gajewski E., Laval J., Dizdaroglu M. Substrate specificity of the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase): excision of purine lesions in DNA produced by ionizing radiation or photosensitization. Biochemistry. 1992 Jan 14;31(1):106–110. doi: 10.1021/bi00116a016. [DOI] [PubMed] [Google Scholar]
  3. Chen J. D., Lacks S. A. Role of uracil-DNA glycosylase in mutation avoidance by Streptococcus pneumoniae. J Bacteriol. 1991 Jan;173(1):283–290. doi: 10.1128/jb.173.1.283-290.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Domena J. D., Timmer R. T., Dicharry S. A., Mosbaugh D. W. Purification and properties of mitochondrial uracil-DNA glycosylase from rat liver. Biochemistry. 1988 Sep 6;27(18):6742–6751. doi: 10.1021/bi00418a015. [DOI] [PubMed] [Google Scholar]
  5. Duncan B. K., Miller J. H. Mutagenic deamination of cytosine residues in DNA. Nature. 1980 Oct 9;287(5782):560–561. doi: 10.1038/287560a0. [DOI] [PubMed] [Google Scholar]
  6. Eftedal I., Guddal P. H., Slupphaug G., Volden G., Krokan H. E. Consensus sequences for good and poor removal of uracil from double stranded DNA by uracil-DNA glycosylase. Nucleic Acids Res. 1993 May 11;21(9):2095–2101. doi: 10.1093/nar/21.9.2095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Impellizzeri K. J., Anderson B., Burgers P. M. The spectrum of spontaneous mutations in a Saccharomyces cerevisiae uracil-DNA-glycosylase mutant limits the function of this enzyme to cytosine deamination repair. J Bacteriol. 1991 Nov;173(21):6807–6810. doi: 10.1128/jb.173.21.6807-6810.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jiricny J., Hughes M., Corman N., Rudkin B. B. A human 200-kDa protein binds selectively to DNA fragments containing G.T mismatches. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8860–8864. doi: 10.1073/pnas.85.23.8860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Krokan H., Wittwer C. U. Uracil DNa-glycosylase from HeLa cells: general properties, substrate specificity and effect of uracil analogs. Nucleic Acids Res. 1981 Jun 11;9(11):2599–2613. doi: 10.1093/nar/9.11.2599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lindahl T. An N-glycosidase from Escherichia coli that releases free uracil from DNA containing deaminated cytosine residues. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3649–3653. doi: 10.1073/pnas.71.9.3649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lindahl T., Ljungquist S., Siegert W., Nyberg B., Sperens B. DNA N-glycosidases: properties of uracil-DNA glycosidase from Escherichia coli. J Biol Chem. 1977 May 25;252(10):3286–3294. [PubMed] [Google Scholar]
  12. Michaels M. L., Tchou J., Grollman A. P., Miller J. H. A repair system for 8-oxo-7,8-dihydrodeoxyguanine. Biochemistry. 1992 Nov 17;31(45):10964–10968. doi: 10.1021/bi00160a004. [DOI] [PubMed] [Google Scholar]
  13. Muller S. J., Caradonna S. Cell cycle regulation of a human cyclin-like gene encoding uracil-DNA glycosylase. J Biol Chem. 1993 Jan 15;268(2):1310–1319. [PubMed] [Google Scholar]
  14. Neddermann P., Jiricny J. The purification of a mismatch-specific thymine-DNA glycosylase from HeLa cells. J Biol Chem. 1993 Oct 5;268(28):21218–21224. [PubMed] [Google Scholar]
  15. Olsen L. C., Aasland R., Wittwer C. U., Krokan H. E., Helland D. E. Molecular cloning of human uracil-DNA glycosylase, a highly conserved DNA repair enzyme. EMBO J. 1989 Oct;8(10):3121–3125. doi: 10.1002/j.1460-2075.1989.tb08464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Selker E. U. Premeiotic instability of repeated sequences in Neurospora crassa. Annu Rev Genet. 1990;24:579–613. doi: 10.1146/annurev.ge.24.120190.003051. [DOI] [PubMed] [Google Scholar]
  17. Shen J. C., Rideout W. M., 3rd, Jones P. A. High frequency mutagenesis by a DNA methyltransferase. Cell. 1992 Dec 24;71(7):1073–1080. doi: 10.1016/s0092-8674(05)80057-1. [DOI] [PubMed] [Google Scholar]
  18. Slupphaug G., Olsen L. C., Helland D., Aasland R., Krokan H. E. Cell cycle regulation and in vitro hybrid arrest analysis of the major human uracil-DNA glycosylase. Nucleic Acids Res. 1991 Oct 11;19(19):5131–5137. doi: 10.1093/nar/19.19.5131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Su S. S., Modrich P. Escherichia coli mutS-encoded protein binds to mismatched DNA base pairs. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5057–5061. doi: 10.1073/pnas.83.14.5057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Talpaert-Borlé M., Clerici L., Campagnari F. Isolation and characterization of a uracil-DNA glycosylase from calf thymus. J Biol Chem. 1979 Jul 25;254(14):6387–6391. [PubMed] [Google Scholar]
  21. Tchou J., Kasai H., Shibutani S., Chung M. H., Laval J., Grollman A. P., Nishimura S. 8-oxoguanine (8-hydroxyguanine) DNA glycosylase and its substrate specificity. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4690–4694. doi: 10.1073/pnas.88.11.4690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tomilin N. V., Aprelikova O. N. Uracil-DNA glycosylases and DNA uracil repair. Int Rev Cytol. 1989;114:125–179. doi: 10.1016/s0074-7696(08)60860-8. [DOI] [PubMed] [Google Scholar]
  23. Vollberg T. M., Siegler K. M., Cool B. L., Sirover M. A. Isolation and characterization of the human uracil DNA glycosylase gene. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8693–8697. doi: 10.1073/pnas.86.22.8693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wallace S. S. AP endonucleases and DNA glycosylases that recognize oxidative DNA damage. Environ Mol Mutagen. 1988;12(4):431–477. doi: 10.1002/em.2860120411. [DOI] [PubMed] [Google Scholar]
  25. Wiebauer K., Jiricny J. In vitro correction of G.T mispairs to G.C pairs in nuclear extracts from human cells. Nature. 1989 May 18;339(6221):234–236. doi: 10.1038/339234a0. [DOI] [PubMed] [Google Scholar]
  26. Wiebauer K., Jiricny J. Mismatch-specific thymine DNA glycosylase and DNA polymerase beta mediate the correction of G.T mispairs in nuclear extracts from human cells. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5842–5845. doi: 10.1073/pnas.87.15.5842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zhang L. H., Vrieling H., van Zeeland A. A., Jenssen D. Spectrum of spontaneously occurring mutations in the hprt gene of V79 Chinese hamster cells. J Mol Biol. 1992 Feb 5;223(3):627–635. doi: 10.1016/0022-2836(92)90979-t. [DOI] [PubMed] [Google Scholar]
  28. de Jong P. J., Grosovsky A. J., Glickman B. W. Spectrum of spontaneous mutation at the APRT locus of Chinese hamster ovary cells: an analysis at the DNA sequence level. Proc Natl Acad Sci U S A. 1988 May;85(10):3499–3503. doi: 10.1073/pnas.85.10.3499. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES