Molecular characterization of the Helicobacter pylori uvrB gene

Stuart A Thompson, Rebecca L. Latch, Martin J. Blaser

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Helicobacter pylori persists in the human stomach where it may encounter a variety of DNA-damaging conditions, including gastric acidity. To determine whether the nucleotide excision repair (NER) pathway contributes to the repair of acid-induced DNA damage, we have cloned the putative H. pylori NER gene, uvrB. Degenerate oligonucleotide primers based on conserved amino acid residues of bacterial UvrB proteins were used in PCR with genomic DNA from H. pylori strain 84-183, and the 1.3-kb PCR product from this reaction was used as a probe to clone uvrB from an H. pylori genomic library. This plasmid clone had a 5.5-kb insert containing a 2.0-kb ORF whose predicted product (658 amino acids; 75.9 kDa) exhibited 69.5% similarity to E. coli UvrB. We constructed an isogenic H. pylori uvrB mutant by inserting a kanamycin-resistance cassette into uvrB and verified its proper placement by Southern hybridization. As with uvrB mutants of other bacteria, the H. pylori uvrB mutant showed a greatly increased sensitivity to the DNA-damaging agents methylmethane sulfonate and ultraviolet radiation. The uvrB mutant also was significantly more sensitive than the wild-type strain to killing by low pH, suggesting that the H. pylori nucleotide excision repair (NER) pathway is involved in the repair of acid-induced DNA damage.

Original languageEnglish (US)
Pages (from-to)113-122
Number of pages10
JournalGene
Volume209
Issue number1-2
DOIs
StatePublished - Mar 16 1998

Fingerprint

Helicobacter pylori
Genes
DNA Repair
DNA Damage
DNA
Stomach
Clone Cells
Kanamycin Resistance
Methyl Methanesulfonate
Amino Acids
Polymerase Chain Reaction
Acids
DNA Primers
Bacterial Proteins
Genomic Library
Open Reading Frames
Plasmids
Radiation
Escherichia coli
Bacteria

Keywords

  • Acid resistance
  • DNA repair
  • Nucleotide excision repair
  • SOS response

ASJC Scopus subject areas

  • Genetics

Cite this

Molecular characterization of the Helicobacter pylori uvrB gene. / Thompson, Stuart A; Latch, Rebecca L.; Blaser, Martin J.

In: Gene, Vol. 209, No. 1-2, 16.03.1998, p. 113-122.

Research output: Contribution to journalArticle

Thompson, Stuart A ; Latch, Rebecca L. ; Blaser, Martin J. / Molecular characterization of the Helicobacter pylori uvrB gene. In: Gene. 1998 ; Vol. 209, No. 1-2. pp. 113-122.
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