Applied Microbiology and Biotechnology

, Volume 98, Issue 10, pp 4625–4636 | Cite as

A SAM-dependent methyltransferase cotranscribed with arsenate reductase alters resistance to peptidyl transferase center-binding antibiotics in Azospirillum brasilense Sp7

Applied microbial and cell physiology
First Online:
Received:
Revised:
Accepted:

Abstract

The genome of Azospirillum brasilense harbors a gene encoding S-adenosylmethionine-dependent methyltransferase, which is located downstream of an arsenate reductase gene. Both genes are cotranscribed and translationally coupled. When they were cloned and expressed individually in an arsenate-sensitive strain of Escherichia coli, arsenate reductase conferred tolerance to arsenate; however, methyltransferase failed to do so. Sequence analysis revealed that methyltransferase was more closely related to a PrmB-type N5-glutamine methyltransferase than to the arsenate detoxifying methyltransferase ArsM. Insertional inactivation of prmB gene in A. brasilense resulted in an increased sensitivity to chloramphenicol and resistance to tiamulin and clindamycin, which are known to bind at the peptidyl transferase center (PTC) in the ribosome. These observations suggested that the inability of prmB:km mutant to methylate L3 protein might alter hydrophobicity in the antibiotic-binding pocket of the PTC, which might affect the binding of chloramphenicol, clindamycin, and tiamulin differentially. This is the first report showing the role of PrmB-type N5-glutamine methyltransferases in conferring resistance to tiamulin and clindamycin in any bacterium.

Keywords

Azospirillum brasilense Arsenate reductase PrmB methyltransferase Chloramphenicol Tiamulin Clindamycin 
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Notes

Acknowledgments

This work was supported by a grant from the Council of Scientific and Industrial Research, New Delhi to AKT. We are grateful to Barry Rosen and Christopher Rensing for providing arsenate-sensitive strains, E. coli WC3110 (DE3) and E. coli AW3110, respectively. We also thank the University Grants Commission, New Delhi and Indian Council of Medical Research, New Delhi for research fellowships to SS and CS.

Supplementary material

253_2014_5574_MOESM1_ESM.pdf (499 kb)
ESM 1 (PDF 498 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Sudhir Singh
    • 1
  • Chhaya Singh
    • 1
  • Anil Kumar Tripathi
    • 1
  1. 1.Laboratory of Bacterial Genetics, School of Biotechnology, Faculty of ScienceBanaras Hindu UniversityVaranasiIndia

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