Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa

Keiko Yokoyama, Yohei Doi, Kunikazu Yamane, Hiroshi Kurokawa, Naohiro Shibata, Keigo Shibayama, Tetsuya Yagi, Haru Kato, Yoshichika Arakawa

Research output: Contribution to journalArticle

170 Citations (Scopus)

Abstract

Background: Bacteria develop resistance to aminoglycosides by producing aminoglycoside-modifying enzymes such as acetyltransferase, phosphorylase, and adenyltransferase. These enzymes, however, cannot confer consistent resistance to various aminoglycosides because of their substrate specificity. Notwithstanding, a Pseudomonas aeruginosa strain AR-2 showing high-level resistance (minimum inhibitory concentration >1024 mg/L) to various aminoglycosides was isolated clinically. We aimed to clone and characterise the genetic determinant of this resistance. Methods: We used conventional methods for DNA manipulation, susceptibility testing, and gene analyses to clone and characterise the genetic determinant of the resistance seen. PCR detection of the gene was also done on a stock of P aeruginosa strains that were isolated clinically since 1997. Findings: An aminoglycoside-resistance gene, designated rmtA, was identified in P aeruginosa AR-2. The Escherichia coli transformant and transconjugant harbouring the rmtA gene showed very high-level resistance to various aminoglycosides, including amikacin, tobramycin, isepamicin, arbekacin, kanamycin, and gentamicin. The 756-bp nucleotide rmtA gene encoded a protein, RmtA. This protein showed considerable similarity to the 16S rRNA methylases of aminoglycoside-producing actinomycetes, which protect bacterial 16S rRNA from intrinsic aminoglycosides by methylation. Incorporation of radiolabelled methyl groups into the 30S ribosome was detected in the presence of RmtA. Of 1113 clinically isolated P aeruginosa strains, nine carried the rmtA gene, as shown by PCR analyses. Interpretation: Our findings strongly suggest intergeneric lateral gene transfer of 16S rRNA methylase gene from some aminoglycoside-producing microorganisms to P aeruginosa. Further dissemination of the rmtA gene in nosocomial bacteria could be a matter of concern in the future.

Original languageEnglish
Pages (from-to)1888-1893
Number of pages6
JournalLancet
Volume362
Issue number9399
DOIs
Publication statusPublished - 06-12-2003
Externally publishedYes

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Aminoglycosides
rRNA Genes
Pseudomonas aeruginosa
Genes
Clone Cells
Bacteria
rRNA (adenosine-O-2'-)methyltransferase
Horizontal Gene Transfer
Polymerase Chain Reaction
Phosphorylases
Acetyltransferases
Tobramycin
Kanamycin
Amikacin
Actinobacteria
Microbial Sensitivity Tests
Enzymes
Substrate Specificity
Gentamicins
Ribosomes

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Yokoyama, K., Doi, Y., Yamane, K., Kurokawa, H., Shibata, N., Shibayama, K., ... Arakawa, Y. (2003). Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa. Lancet, 362(9399), 1888-1893. https://doi.org/10.1016/S0140-6736(03)14959-8
Yokoyama, Keiko ; Doi, Yohei ; Yamane, Kunikazu ; Kurokawa, Hiroshi ; Shibata, Naohiro ; Shibayama, Keigo ; Yagi, Tetsuya ; Kato, Haru ; Arakawa, Yoshichika. / Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa. In: Lancet. 2003 ; Vol. 362, No. 9399. pp. 1888-1893.
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Yokoyama, K, Doi, Y, Yamane, K, Kurokawa, H, Shibata, N, Shibayama, K, Yagi, T, Kato, H & Arakawa, Y 2003, 'Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa', Lancet, vol. 362, no. 9399, pp. 1888-1893. https://doi.org/10.1016/S0140-6736(03)14959-8

Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa. / Yokoyama, Keiko; Doi, Yohei; Yamane, Kunikazu; Kurokawa, Hiroshi; Shibata, Naohiro; Shibayama, Keigo; Yagi, Tetsuya; Kato, Haru; Arakawa, Yoshichika.

In: Lancet, Vol. 362, No. 9399, 06.12.2003, p. 1888-1893.

Research output: Contribution to journalArticle

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T1 - Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa

AU - Yokoyama, Keiko

AU - Doi, Yohei

AU - Yamane, Kunikazu

AU - Kurokawa, Hiroshi

AU - Shibata, Naohiro

AU - Shibayama, Keigo

AU - Yagi, Tetsuya

AU - Kato, Haru

AU - Arakawa, Yoshichika

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N2 - Background: Bacteria develop resistance to aminoglycosides by producing aminoglycoside-modifying enzymes such as acetyltransferase, phosphorylase, and adenyltransferase. These enzymes, however, cannot confer consistent resistance to various aminoglycosides because of their substrate specificity. Notwithstanding, a Pseudomonas aeruginosa strain AR-2 showing high-level resistance (minimum inhibitory concentration >1024 mg/L) to various aminoglycosides was isolated clinically. We aimed to clone and characterise the genetic determinant of this resistance. Methods: We used conventional methods for DNA manipulation, susceptibility testing, and gene analyses to clone and characterise the genetic determinant of the resistance seen. PCR detection of the gene was also done on a stock of P aeruginosa strains that were isolated clinically since 1997. Findings: An aminoglycoside-resistance gene, designated rmtA, was identified in P aeruginosa AR-2. The Escherichia coli transformant and transconjugant harbouring the rmtA gene showed very high-level resistance to various aminoglycosides, including amikacin, tobramycin, isepamicin, arbekacin, kanamycin, and gentamicin. The 756-bp nucleotide rmtA gene encoded a protein, RmtA. This protein showed considerable similarity to the 16S rRNA methylases of aminoglycoside-producing actinomycetes, which protect bacterial 16S rRNA from intrinsic aminoglycosides by methylation. Incorporation of radiolabelled methyl groups into the 30S ribosome was detected in the presence of RmtA. Of 1113 clinically isolated P aeruginosa strains, nine carried the rmtA gene, as shown by PCR analyses. Interpretation: Our findings strongly suggest intergeneric lateral gene transfer of 16S rRNA methylase gene from some aminoglycoside-producing microorganisms to P aeruginosa. Further dissemination of the rmtA gene in nosocomial bacteria could be a matter of concern in the future.

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Yokoyama K, Doi Y, Yamane K, Kurokawa H, Shibata N, Shibayama K et al. Acquisition of 16S rRNA methylase gene in Pseudomonas aeruginosa. Lancet. 2003 Dec 6;362(9399):1888-1893. https://doi.org/10.1016/S0140-6736(03)14959-8