Home LiteratureArticle Details
PMID: 1886771 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Human uracil-DNA glycosylase complements E. coli ung mutants.

Nucleic acids research ·Vol. 19 ·No. 16 ·1991-08-25 ·Pages 4473-8

Olsen LC, Aasland R, Krokan HE, Helland DE

Abstract

We have previously isolated a cDNA encoding a human uracil-DNA glycosylase which is closely related to the bacterial and yeast enzymes. In vitro expression of this cDNA produced a protein with an apparent molecular weight of 34 K in agreement with the size predicted from the sequence data. The in vitro expressed protein exhibited uracil-DNA glycosylase activity. The close resemblance between the human and the bacterial enzyme raised the possibility that the human enzyme may be able to complement E. coli ung mutants. In order to test this hypothesis, the human uracil-DNA glycosylase cDNA was established in a bacterial expression vector. Expression of the human enzyme as a LacZ alpha-humUNG fusion protein was then studied in E. coli ung mutants. E. coli cells lacking uracil-DNA glycosylase activity exhibit a weak mutator phenotype and they are permissive for growth of phages with uracil-containing DNA. Here we show that the expression of human uracil-DNA glycosylase in E. coli can restore the wild type phenotype of ung mutants. These results demonstrate that the evolutionary conservation of the uracil-DNA glycosylase structure is also reflected in the conservation of the mechanism for removal of uracil from DNA.

MeSH Terms
Amino Acid Sequence Base Sequence DNA Glycosylases DNA, Bacterial/metabolism Electrophoresis, Polyacrylamide Gel Escherichia coli/enzymology,metabolism Gene Expression/physiology Genetic Complementation Test Humans Molecular Sequence Data N-Glycosyl Hydrolases/genetics,metabolism Recombinant Fusion Proteins/biosynthesis Transformation, Genetic Uracil/metabolism Uracil-DNA Glycosidase
Chemicals
DNA, Bacterial Recombinant Fusion Proteins Uracil DNA Glycosylases N-Glycosyl Hydrolases Uracil-DNA Glycosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Olsen L C
Laboratory of Biotechnology, University of Bergen, Norway.
Aasland R
Krokan H E
Helland D E
References (36)
36 references, click to expand
  1. Heat-induced deamination of cytosine residues in deoxyribonucleic acid.
    Biochemistry. 1974 Jul 30;13(16):3405-10 PMID: 4601435
  2. 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):3649-53 PMID: 4610583
  3. Isolation and structure of a cDNA expressing a mammalian 3-methyladenine-DNA glycosylase.
    EMBO J. 1990 Oct;9(10):3337-42 PMID: 1698614
  4. Isolation and characterization of a human cDNA encoding uracil-DNA glycosylase.
    Biochim Biophys Acta. 1991 Feb 16;1088(2):197-207 PMID: 2001396
  5. Nucleotide sequence of the Streptococcus pneumoniae ung gene encoding uracil-DNA glycosylase.
    Nucleic Acids Res. 1990 Nov 25;18(22):6693 PMID: 2251133
  6. Rat DNA polymerase beta gene can join in excision repair of Escherichia coli.
    Nucleic Acids Res. 1990 Oct 11;18(19):5673-6 PMID: 2216761
  7. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
    Science. 1988 Jan 29;239(4839):487-91 PMID: 2448875
  8. Molecular cloning of human uracil-DNA glycosylase, a highly conserved DNA repair enzyme.
    EMBO J. 1989 Oct;8(10):3121-5 PMID: 2555154
  9. Sequence analysis, expression, and conservation of Escherichia coli uracil DNA glycosylase and its gene (ung).
    J Biol Chem. 1988 Jun 5;263(16):7776-84 PMID: 2836397
  10. Identification of the coding sequence for herpes simplex virus uracil-DNA glycosylase.
    J Virol. 1988 Dec;62(12):4774-7 PMID: 2846888
  11. High efficiency transformation of E. coli by high voltage electroporation.
    Nucleic Acids Res. 1988 Jul 11;16(13):6127-45 PMID: 3041370
  12. Gene UL2 of herpes simplex virus type 1 encodes a uracil-DNA glycosylase.
    J Gen Virol. 1989 Feb;70 ( Pt 2):449-54 PMID: 2567340
  13. Uracil-DNA glycosylase inhibitor gene of bacteriophage PBS2 encodes a binding protein specific for uracil-DNA glycosylase.
    J Biol Chem. 1989 Jan 15;264(2):1163-71 PMID: 2492016
  14. Molecular cloning and primary structure of the uracil-DNA-glycosylase gene from Saccharomyces cerevisiae.
    J Biol Chem. 1989 Feb 15;264(5):2593-8 PMID: 2644266
  15. Isolation and characterization of the human uracil DNA glycosylase gene.
    Proc Natl Acad Sci U S A. 1989 Nov;86(22):8693-7 PMID: 2813420
  16. Purification and determination of the NH2-terminal amino acid sequence of uracil-DNA glycosylase from human placenta.
    Biochemistry. 1989 Jan 24;28(2):780-4 PMID: 2713345
  17. Purification and properties of mitochondrial uracil-DNA glycosylase from rat liver.
    Biochemistry. 1988 Sep 6;27(18):6742-51 PMID: 3196681
  18. Physical association of the human base-excision repair enzyme uracil DNA glycosylase with the 70,000-dalton catalytic subunit of DNA polymerase alpha.
    Proc Natl Acad Sci U S A. 1986 Oct;83(20):7608-12 PMID: 3463989
  19. Purification and properties of the human placental uracil DNA glycosylase.
    Biochim Biophys Acta. 1987 Aug 13;925(2):226-33 PMID: 3620497
  20. Purification of nuclear and mitochondrial uracil-DNA glycosylase from rat liver. Identification of two distinct subcellular forms.
    Biochemistry. 1985 Dec 3;24(25):7320-8 PMID: 3910103
  21. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92 PMID: 3881765
  22. Uracil-DNA glycosylase in HeLa S3 cells: interconvertibility of 50 and 20 kDa forms and similarity of the nuclear and mitochondrial form of the enzyme.
    Biochim Biophys Acta. 1985 Dec 20;832(3):308-18 PMID: 4074751
  23. Sequential initiation of lagging and leading strand synthesis by two different polymerase complexes at the SV40 DNA replication origin.
    Nature. 1990 Aug 9;346(6284):534-9 PMID: 2165567
  24. Isolation and structural characterization of a cDNA clone encoding the human DNA repair protein for O6-alkylguanine.
    Proc Natl Acad Sci U S A. 1990 Jan;87(2):686-90 PMID: 2405387
  25. The enzymology of apurinic/apyrimidinic endonucleases.
    Mutat Res. 1990 Sep-Nov;236(2-3):173-201 PMID: 1697933
  26. Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169.
    Curr Top Microbiol Immunol. 1990;154:125-69 PMID: 2161319
  27. Chromosomal assignment of human uracil-DNA glycosylase to chromosome 12.
    Genomics. 1990 May;7(1):139-41 PMID: 2335354
  28. The presence of nuclear and mitochondrial uracil-DNA glycosylase in extracts of human KB cells.
    Nucleic Acids Res. 1980 Feb 25;8(4):875-88 PMID: 6253928
  29. Specificity of the bacteriophage PBS2 induced inhibitor of uracil-DNA glycosylase.
    Biochemistry. 1981 Oct 13;20(21):6092-6 PMID: 6796110
  30. Uracil DNA-glycosylase. Purification and properties of this enzyme isolated from blast cells of acute myelocytic leukemia patients.
    J Biol Chem. 1980 Mar 25;255(6):2293-300 PMID: 6766936
  31. Uracil DNa-glycosylase from HeLa cells: general properties, substrate specificity and effect of uracil analogs.
    Nucleic Acids Res. 1981 Jun 11;9(11):2599-613 PMID: 7279657
  32. DNA glycosylases, endonucleases for apurinic/apyrimidinic sites, and base excision-repair.
    Prog Nucleic Acid Res Mol Biol. 1979;22:135-92 PMID: 392601
  33. The properties of a bacteriophage T5 mutant unable to induce deoxyuridine 5'-triphosphate nucleotidohydrolase. Synthesis of uracil-containing T5 deoxyribonucleic acid.
    J Biol Chem. 1979 Aug 25;254(16):7534-9 PMID: 381286
  34. Escherichia coli K-12 mutants deficient in uracil-DNA glycosylase.
    J Bacteriol. 1978 Jun;134(3):1039-45 PMID: 350837
  35. DNA N-glycosidases: properties of uracil-DNA glycosidase from Escherichia coli.
    J Biol Chem. 1977 May 25;252(10):3286-94 PMID: 324994
  36. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/19.16.4473
Published
1991-08-25
Pages
4473-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC328636
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/19.16.4473 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2