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Journal of Bacteriology, October 2007, p. 6919-6927, Vol. 189, No. 19
0021-9193/07/$08.00+0     doi:10.1128/JB.00904-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Regulatory Overlap and Functional Redundancy among Bacillus subtilis Extracytoplasmic Function {sigma} Factors{triangledown}

Thorsten Mascher,{dagger} Anna-Barbara Hachmann, and John D. Helmann*

Department of Microbiology, Cornell University, Ithaca, New York 14853-8101

Received 8 June 2007/ Accepted 23 July 2007

Bacillus subtilis encodes seven extracytoplasmic function (ECF) {sigma} factors that regulate partially overlapping regulons related to cell envelope homeostasis and antibiotic resistance. Here, we investigated their physiological role by constructing a mutant set of single, double, triple, and quadruple ECF {sigma} factor deletions in the undomesticated B. subtilis strain NCIB3610. This mutant set was subsequently screened for defects in motility, multicellular differentiation, and sensitivity to more than 200 chemicals by using Phenotype MicroArrays. A quadruple mutant strain, harboring deletions of the sigV, sigY, sigZ, and ylaC gene, behaved indistinguishably from the wild-type strain, indicative of either regulatory redundancy or very specific functions of these four ECF {sigma} factors. In contrast, a triple mutant, inactivated for the sigM, sigW, and sigX genes (but none of the corresponding double mutants), showed a biphasic growth behavior and a complete loss of multicellular differentiation, as judged by both colony formation and the inability to form a pellicle. This triple mutant also displayed a greatly increased sensitivity to detergents and several cell wall antibiotics including ß-lactams, polymyxin B, and D-cycloserine. In several cases, these antibiotic-sensitive phenotypes are significantly enhanced in the triple mutant strain relative to strains lacking only one or two {sigma} factors.

* Corresponding author: Department of Microbiology, Wing Hall, Cornell University, Ithaca, NY 14853-8101. Phone: (607) 255-6570. Fax: (607) 255-3904. E-mail: jdh9{at}cornell.edu

{triangledown} Published ahead of print on 3 August 2007.

{dagger} Present address: Department of General Microbiology, Institute of Microbiology and Genetics, Georg-August-University, Grisebachstr. 8, D-37077 Göttingen, Germany.

Journal of Bacteriology, October 2007, p. 6919-6927, Vol. 189, No. 19
0021-9193/07/$08.00+0     doi:10.1128/JB.00904-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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