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Growth, osmotic downshock resistance and differentiation of Bacillus subtilis strains lacking mechanosensitive channels

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Abstract

Previous work has shown that the mechanosensitive (MS) channel of large conductance (MscL) is essential for preventing lysis of Bacillus subtilis log phase cells upon a rapid, severe osmotic downshock. Growing cells of B. subtilis strains lacking MscL and one or more putative MS channel proteins of small conductance (YhdY, YkuT and YfkC) showed even higher sensitivity to an osmotic downshock. The effect was greatest for a strain lacking MscL and YkuT, and a strain lacking all four MS channel proteins had a similar phenotype. These defects were complemented by expression of either MscL or YkuT in trans. All MS channel mutant strains ultimately became resistant to osmotic downshock in stationary phase but at varying times, with mscL ykuT strains taking the longest time to become resistant. Expression of β-galactosidase from gene fusions to lacZ showed modest expression of ykuT and lower levels of expression of yhdY and yfkC when strains were grown in medium containing high salt. Sporulation of all MS channel mutant strains was normal, and the mutant spores germinated normally with l-alanine or dodecylamine.

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Abbreviations

DPA:

Pyridine-2-6-dicarboxylic acid or dipicolinic acid

LB:

Luria-Bertani medium

MS:

Mechanosensitive

MscL:

Mechanosensitive channel of large conductance

MscS:

Mechanosensitive channel of small conductance

OD600 nm :

Optical density at 600 nm

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Acknowledgments

This work was supported by a grant from the Army Research Office. We are grateful to Fabrizio Arigoni for supplying plasmid pRDC18.

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Authors and Affiliations

  1. Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030-3305, USA

    Paul G. Wahome & Peter Setlow

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  1. Paul G. Wahome
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  2. Peter Setlow
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Correspondence to Peter Setlow.

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Communicated by Erko Stackebrandt.

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Wahome, P.G., Setlow, P. Growth, osmotic downshock resistance and differentiation of Bacillus subtilis strains lacking mechanosensitive channels. Arch Microbiol 189, 49–58 (2008). https://doi.org/10.1007/s00203-007-0292-z

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  • DOI: https://doi.org/10.1007/s00203-007-0292-z

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