Overexpression of spoT gene in coccoid forms of clinical Helicobacter pylori isolates

Folia Microbiol (Praha). 2018 Jul;63(4):459-465. doi: 10.1007/s12223-017-0557-0. Epub 2018 Jan 11.

Abstract

Helicobacter pylori (H. pylori) can convert to coccoid form in unfavorable conditions or as a result of antibiotic treatment. In order to adapt to harsh environments, H. pylori requires a stringent response which, encoded by the spoT gene, has a bifunctional enzyme possessing both (p)ppGpp synthetic and degrading activity. Our goal in this study was to compare spoT gene expression in spiral and induced coccoid forms of H. pylori with use of amoxicillin. First, clinical isolate coccoid forms were induced with amoxicillin; then, the viability test was analyzed by flow cytometer. After RNA extraction, cDNA synthesis and designing a specific primer for spoT gene, evaluation of the desired gene expression in both forms were studied. Bacterial isolates exposed to amoxicillin at MIC and 1/2 MIC induced morphological conversion better and faster than other MIC concentration. The expression of spoT gene was significantly downregulated in spiral forms of H. pylori, while the gene expression was upregulated and + 30.3-fold changes was seen in coccoid forms of bacterium. To summarize, spoT gene is one of the key factors for antibiotic resistance and its enhanced expression in coccoid form can be a valuable diagnostic marker for recognition of H. pylori during morphological conversion.

MeSH terms

  • Adaptation, Physiological / genetics
  • Amoxicillin / metabolism*
  • Amoxicillin / pharmacology
  • Anti-Bacterial Agents / metabolism*
  • Anti-Bacterial Agents / pharmacology
  • Drug Resistance, Bacterial / genetics
  • Gene Expression Regulation, Bacterial / drug effects
  • Gene Expression Regulation, Bacterial / physiology*
  • Genes, Bacterial / genetics
  • Helicobacter pylori / drug effects
  • Helicobacter pylori / genetics*
  • Helicobacter pylori / growth & development*
  • Microbial Sensitivity Tests
  • Microbial Viability / drug effects
  • Pyrophosphatases / genetics

Substances

  • Anti-Bacterial Agents
  • Amoxicillin
  • Pyrophosphatases