Increased resistance against tellurite is conferred by a mutation in the promoter region of uncommon tellurite resistance gene tehB in the ter -negative Shiga toxin-producing Escherichia coli O157:H7

Author:

Matsumoto Yuko1,Lee Kenichi2ORCID,Akasaka Ryuya23,Honjo Hayato23,Koizumi Mitsumasa1,Sato Toshio4,Kubomura Akiko2ORCID,Ishijima Nozomi2,Akeda Yukihiro2,Ohnishi Makoto2,Iyoda Sunao2

Affiliation:

1. Yokohama Institute of Public Health, Kanagawa, Japan

2. Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan

3. Tokyo College of Biotechnology, Tokyo, Japan

4. Japan Microbiological Laboratory Co. Ltd., Miyagi, Japan

Abstract

ABSTRACT Resistance to potassium tellurite (PT) is an important indicator in isolating Shiga toxin-producing Escherichia coli (STEC) O157:H7 and other major STEC serogroups. Common resistance determinant genes are encoded in the ter gene cluster. We found an O157:H7 isolate that does not harbor ter but is resistant to PT. One nonsynonymous mutation was found in another PT resistance gene, tehA , through whole-genome sequence analyses. To elucidate the contribution of this mutation to PT resistance, complementation of tehA and the related gene tehB in isogenic strains and quantitative RT‒PCR were performed. The results indicated that the point mutation not only changed an amino acid of tehA , but also was positioned on a putative internal promoter of tehB and increased PT resistance by elevating tehB mRNA expression. Meanwhile, the amino acid change in tehA had negligible impact on the PT resistance. Comprehensive screening revealed that 2.3% of O157:H7 isolates in Japan did not harbor the ter gene cluster, but the same mutation in tehA was not found. These results suggested that PT resistance in E. coli can be enhanced through one mutational event even in ter -negative strains. IMPORTANCE Selective agents are important for isolating Shiga toxin-producing Escherichia coli (STEC) because the undesirable growth of microflora should be inhibited. Potassium tellurite (PT) is a common selective agent for major STEC serotypes. In this study, we found a novel variant of PT resistance genes, tehAB , in STEC O157:H7. Molecular experiments clearly showed that one point mutation in a predicted internal promoter region of tehB upregulated the expression of the gene and consequently led to increased resistance to PT. Because tehAB genes are ubiquitous across E. coli , these results provide universal insight into PT resistance in this species.

Funder

Japan Agency for Medical Research and Development

MEXT | Japan Society for the Promotion of Science

Ministry of Health, Labour and Welfare

Publisher

American Society for Microbiology

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