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Mutations in the gyrB, parC, and parE Genes of Quinolone-Resistant Isolates and Mutants of Edwardsiella tarda

  • Kim, Myoung-Sug (Pathology Division, National Fisheries Research and Development Institute) ;
  • Jun, Lyu-Jin (Faculty of Applied Marine Science, College of Ocean Science, Jeju National University) ;
  • Shin, Soon-Bum (Food and Safety Research Center, National Fisheries Research and Development Institute) ;
  • Park, Myoung-Ae (Pathology Division, National Fisheries Research and Development Institute) ;
  • Jung, Sung-Hee (Pathology Division, National Fisheries Research and Development Institute) ;
  • Kim, Kwang-Il (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Moon, Kyung-Ho (College of Pharmacy, Kyungsung University) ;
  • Jeong, Hyun-Do (Department of Aquatic Life Medicine, Pukyong National University)
  • 투고 : 2010.09.06
  • 심사 : 2010.10.19
  • 발행 : 2010.12.28

초록

The full-length genes gyrB (2,415 bp), parC (2,277 bp), and parE (1,896 bp) in Edwardsiella tarda were cloned by PCR with degenerate primers based on the sequence of the respective quinolone resistance-determining region (QRDR), followed by elongation of 5' and 3' ends using cassette ligation-mediated PCR (CLMP). Analysis of the cloned genes revealed open reading frames (ORFs) encoding proteins of 804 (GyrB), 758 (ParC), and 631 (ParE) amino acids with conserved gyrase/topoisomerase features and motifs important for enzymatic function. The ORFs were preceded by putative promoters, ribosome binding sites, and inverted repeats with the potential to form cruciform structures for binding of DNA-binding proteins. When comparing the deduced amino acid sequences of E. tarda GyrB, ParC, and ParE with those of the corresponding proteins in other bacteria, they were found to be most closely related to Escherichia coli GyrB (87.6% identity), Klebsiella pneumoniae ParC (78.8% identity), and Salmonella Typhimurium ParE (89.5% identity), respectively. The two topoisomerase genes, parC and parE, were found to be contiguous on the E. tarda chromosome. All 18 quinolone-resistant isolates obtained from Korea thus far did not contain subunit alternations apart from a substitution in GyrA (Ser83$\rightarrow$Arg). However, an alteration in the QRDR of ParC (Ser84$\rightarrow$Ile) following an amino acid substitution in GyrA (Asp87$\rightarrow$Gly) was detected in E. tarda mutants selected in vitro at $8{\mu}g/ml$ ciprofloxacin (CIP). A mutant with a GyrB (Ser464$\rightarrow$Leu) and GyrA (Asp87$\rightarrow$Gly) substitution did not show a significant increase in the minimum inhibitory concentration (MIC) of CIP. None of the in vitro mutants exhibited mutations in parE. Thus, gyrA and parC should be considered to be the primary and secondary targets, respectively, of quinolones in E. tarda.

키워드

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피인용 문헌

  1. DNA Gyrase and Topoisomerase IV Mutations and their effect on Quinolones Resistant Proteus mirabilis among UTIs Patients vol.36, pp.6, 2020, https://doi.org/10.12669/pjms.36.6.2207