Cloning and characterization of uracil-DNA glycosylase and the biological consequences of the loss of its function in the nematode Caenorhabditis elegans

Nobuya Nakamura, Hironobu Morinaga, Masahiro Kikuchi, Shinichirou Yonekura, Naoaki Ishii, Kazuo Yamamoto, Shuji Yonei, Qiu Mei Zhang

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Uracil arises in DNA from spontaneous deamination of cytosine and through incorporation of dUMP by DNA polymerase during DNA replication. Excision of uracil by the action of uracil-DNA glycosylase (Ung) initiates the base excision repair pathway to counter the promutagenic base modification. In this study, we cloned a cDNA-encoding Caenorhabditis elegans homologue (CeUng-1) of Escherichia coli Ung. There was 49% identity in amino acid sequence between E.coli Ung and CeUng-1. Purified CeUng-1 removed uracil from both U:G and U:A base pairs in DNA. It also removed uracil from single-stranded oligonucleotide substrate less efficiently than double-stranded oligonucleotide. The CeUng-1 activity was inhibited by Bacillus subtilis Ung inhibitor, indicating that CeUng-1 is a member of the family-1 Ung group. The mutation in the ung-1 gene did not affect development, fertility and lifespan in C.elegans, suggesting the existence of backup enzyme. However, we could not detect residual uracil excision activity in the extract derived from the ung-1 mutant. The present experiments also showed that the ung-1 mutant of C.elegans was more resistant to NaHSO3-inducing cytosine deamination than wild-type strain.

Original languageEnglish
Pages (from-to)407-413
Number of pages7
JournalMutagenesis
Volume23
Issue number5
DOIs
Publication statusPublished - Sep 2008
Externally publishedYes

Fingerprint

Uracil-DNA Glycosidase
Uracil
Cloning
Caenorhabditis elegans
Organism Cloning
Deamination
Cytosine
Oligonucleotides
Escherichia coli
DNA
Bacilli
DNA-Directed DNA Polymerase
Bacillus subtilis
DNA Replication
Base Pairing
DNA Repair
Fertility
Amino Acid Sequence
Repair
Complementary DNA

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Cloning and characterization of uracil-DNA glycosylase and the biological consequences of the loss of its function in the nematode Caenorhabditis elegans. / Nakamura, Nobuya; Morinaga, Hironobu; Kikuchi, Masahiro; Yonekura, Shinichirou; Ishii, Naoaki; Yamamoto, Kazuo; Yonei, Shuji; Zhang, Qiu Mei.

In: Mutagenesis, Vol. 23, No. 5, 09.2008, p. 407-413.

Research output: Contribution to journalArticle

Nakamura, Nobuya ; Morinaga, Hironobu ; Kikuchi, Masahiro ; Yonekura, Shinichirou ; Ishii, Naoaki ; Yamamoto, Kazuo ; Yonei, Shuji ; Zhang, Qiu Mei. / Cloning and characterization of uracil-DNA glycosylase and the biological consequences of the loss of its function in the nematode Caenorhabditis elegans. In: Mutagenesis. 2008 ; Vol. 23, No. 5. pp. 407-413.
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