Journal
MOLECULAR BIOSYSTEMS
Volume 8, Issue 6, Pages 1806-1814Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c2mb25031d
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Funding
- German Federal Ministry of Education and Research (BMBF) [BaCell-(0313978)]
- BaCell2 project [0315784]
- Cosmic2 project [0315782D]
- SysMO project
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In Bacillus subtilis the sigma(B) mediated general stress response provides protection against various environmental and energy related stress conditions. To better understand the general stress response, we need to explore the mechanism by which the components interact. Here, we performed experiments in B. subtilis wild type and mutant strains to test and validate a mathematical model of the dynamics of sigma(B) activity. In the mutant strain BSA115, sigma(B) transcription is inducible by the addition of IPTG and negative control of sigma(B) activity by the anti-sigma factor RsbW is absent. In contrast to our expectations of a continuous beta-galactosidase activity from a ctc::lacZ fusion, we observed a transient activity in the mutant. To explain this experimental finding, we constructed mathematical models reflecting different hypotheses regarding the regulation of sigma(B) and beta-galactosidase dynamics. Only the model assuming instability of either ctc:: lacZ mRNA or beta-galactosidase protein is able to reproduce the experiments in silico. Subsequent Northern blot experiments revealed stable high-level ctc:: lacZ mRNA concentrations after the induction of the sigma(B) response. Therefore, we conclude that protein instability following sigma(B) activation is the most likely explanation for the experimental observations. Our results thus support the idea that B. subtilis increases the cytoplasmic proteolytic degradation to adapt the proteome in face of environmental challenges following activation of the general stress response. The findings also have practical implications for the analysis of stress response dynamics using lacZ reporter gene fusions, a frequently used strategy for the sigma(B) response.
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