NDLI: Proteolysis of beta-galactosidase following SigmaB activation in Bacillus subtilis

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Proteoglycan sequence

Genome-scale genetic manipulation methods for exploring bacterial molecular biology

Endothelial cell's biomechanical properties are regulated by invasive cancer cells

Synthetic antibodies as tools to probe RNA-binding protein function

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Integrated investigation of lipidome and related signaling pathways uncovers molecular mechanisms of tetramethylpyrazine and butylidenephthalide protecting endothelial cells under oxidative stress

Backbone conformational preferences of an intrinsically disordered protein in solution

Proteolysis of beta-galactosidase following SigmaB activation in Bacillus subtilis

Engineering a large protein by combined rational and random approaches: stabilizing the Clostridium thermocellum cellobiose phosphorylase

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Proteolysis of beta-galactosidase following SigmaB activation in Bacillus subtilis

Content Provider	Royal Society of Chemistry (RSC)
Author	Millat, Thomas Liebal, Ulf W. Steil, Leif Homuth, Georg Wolkenhauer, Olaf Völker, Uwe Sappa, Praveen K.
Copyright Year	2012
Abstract	In Bacillus subtilis the σB 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 σB activity. In the mutant strain BSA115, σB transcription is inducible by the addition of IPTG and negative control of σB activity by the anti-sigma factor RsbW is absent. In contrast to our expectations of a continuous β-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 σB and β-galactosidase dynamics. Only the model assuming instability of either ctc::lacZ mRNA or β-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 σB response. Therefore, we conclude that protein instability following σB 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 σB response.
Starting Page	1806
Ending Page	1814
Page Count	9
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	8
Issue Number	6
Journal	Molecular BioSystems
DOI	10.1039/c2mb25031d
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Beta-galactosidase Lac operon Isopropyl β-D-1-thiogalactopyranoside Proteome Protein Northern blot Messenger RNA Reporter gene Gene Bacillus IPTG Gene regulatory network Mutation Mathematical model
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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Different Mechanisms for Thermal Inactivation of Bacillus subtilis Signal Peptidase Mutants

Light-responsive control of bacterial gene expression: precise triggering of the lac promoter activity using photocaged IPTG

Regulated expression of foreign genes in vaccinia virus under the control of bacteriophage t7 rna polymerase and the escherichia coli lac repressor (1991).

Suitability of different β-galactosidases as reporter enzymes in Bacillus subtilis

Cloning and expression of dnaK gene from Bacillus pumilus of hot water spring origin.

Multiple pathways of Spx (YjbD) proteolysis in Bacillus subtilis.

Catabolite repression of the lac operon. Separt epressionof two enzymes.

Functional Analysis of Paralogous Thiol-disulfide Oxidoreductases in Bacillus subtilis

Nucleotide sequence of the sporulation operon, spoiiie, of bacillus subtilis (1986).

Proteolysis of beta-galactosidase following SigmaB activation in Bacillus subtilis

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Different Mechanisms for Thermal Inactivation of Bacillus subtilis Signal Peptidase Mutants

Light-responsive control of bacterial gene expression: precise triggering of the lac promoter activity using photocaged IPTG

Regulated expression of foreign genes in vaccinia virus under the control of bacteriophage t7 rna polymerase and the escherichia coli lac repressor (1991).

Suitability of different β-galactosidases as reporter enzymes in Bacillus subtilis

Cloning and expression of dnaK gene from Bacillus pumilus of hot water spring origin.

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Proteolysis of beta-galactosidase following SigmaB activation in Bacillus subtilis