Osmoregulation of the <i>opuE</i> proline transport gene from <i>Bacillus subtilis</i>: contributions of the sigma A‐ and sigma B‐dependent stress‐responsive promoters

Spiegelhalter, Frank; Bremer, Erhard

doi:10.1046/j.1365-2958.1998.00929.x

Cited by 71 publications

(109 citation statements)

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“…Strain JSB17 [proA6pEPV1T (tet)] was isolated after transformation of strain JH642 with the circular plasmid pEPV1T (Ogura et al, 1994) and selection for tetracycline-resistant colonies; in this strain, plasmid pEPV1T is inserted into the proA gene via a Campbell-type single-crossover integration event, thereby disrupting the integrity of proA. The proA mutation present in strain JSB17 was introduced into strain FSB1 [(treA : : neo)1] (Spiegelhalter & Bremer, 1998) by DNA transformation with chromosomal DNA from strain JSB17 and subsequent selection for tetracycline-resistant colonies. This yielded strain JSB34 [D(treA : : neo) (proA6pEPV1T (tet)].…”

Section: Methodsmentioning

confidence: 99%

“…Proline also serves as an important osmostress protectant (Bremer, 2002). For this task, proline can either be taken up by B. subtilis from the environment via the osmotically inducible OpuE transporter (Spiegelhalter & Bremer, 1998;von Blohn et al, 1997) or be produced in very large quantities from the precursor glutamate (Belitsky et al, 2001;Whatmore et al, 1990). Proline belongs to a selected group of organic osmolytes, the compatible solutes, which are amassed by many micro-organisms to adapt to high-osmolarity growth conditions (Kempf & Bremer, 1998).…”

Section: Introductionmentioning

confidence: 99%

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T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis

et al. 2011

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Bacillus subtilis possesses interlinked routes for the synthesis of proline. The ProJ-ProA-ProH route is responsible for the production of proline as an osmoprotectant, and the ProB-ProA-ProI route provides proline for protein synthesis. We show here that the transcription of the anabolic proBA and proI genes is controlled in response to proline limitation via a T-box-mediated termination/antitermination regulatory mechanism, a tRNA-responsive riboswitch. Primer extension analysis revealed mRNA leader transcripts of 270 and 269 nt for the proBA and proI genes, respectively, both of which are synthesized from SigA-type promoters. These leader transcripts are predicted to fold into two mutually exclusive secondary mRNA structures, forming either a terminator or an antiterminator configuration. Northern blot analysis allowed the detection of both the leader and the full-length proBA and proI transcripts. Assessment of the level of the proBA transcripts revealed that the amount of the full-length mRNA species strongly increased in proline-starved cultures. Genetic studies with a proB-treA operon fusion reporter strain demonstrated that proBA transcription is sensitively tied to proline availability and is derepressed as soon as cellular starvation for proline sets in. Both the proBA and the proI leader sequences contain a CCU proline-specific specifier codon prone to interact with the corresponding uncharged proline-specific tRNA. By replacing the CCU proline specifier codon in the proBA T-box leader with UUC, a codon recognized by a Phe-specific tRNA, we were able to synthetically re-engineer the proline-specific control of proBA transcription to a control that was responsive to starvation for phenylalanine.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis

et al. 2011

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“…opuE expression is controlled from σ B and σ A promoters, both of which are activated under salt stress (41). The presence of the σ B -independent regulatory pathway should make activation of OpuE under salt stress less rate responsive than a pure σ B -dependent target.…”

Section: Colony Promoter Activity (Au)mentioning

confidence: 99%

Rate of environmental change determines stress response specificity

Young

Locke

Elowitz

2013

Proc. Natl. Acad. Sci. U.S.A.

116

143

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Cells use general stress response pathways to activate diverse target genes in response to a variety of stresses. However, general stress responses coexist with more specific pathways that are activated by individual stresses, provoking the fundamental question of whether and how cells control the generality or specificity of their response to a particular stress. Here we address this issue using quantitative time-lapse microscopy of the Bacillus subtilis environmental stress response, mediated by σ B . We analyzed σ B activation in response to stresses such as salt and ethanol imposed at varying rates of increase. Dynamically, σ B responded to these stresses with a single adaptive activity pulse, whose amplitude depended on the rate at which the stress increased. This rate-responsive behavior can be understood from mathematical modeling of a key negative feedback loop in the underlying regulatory circuit. Using RNAseq we analyzed the effects of both rapid and gradual increases of ethanol and salt stress across the genome. Because of the rate responsiveness of σ B activation, salt and ethanol regulons overlap under rapid, but not gradual, increases in stress. Thus, the cell responds specifically to individual stresses that appear gradually, while using σ B to broaden the cellular response under more rapidly deteriorating conditions. Such dynamic control of specificity could be a critical function of other general stress response pathways.systems biology | single-cell dynamics | computational biology C ells must respond to, and anticipate, a wide range of stresses that occur on multiple timescales. For this purpose, many species use general stress response pathways, which activate a diverse set of target regulons in response to a variety of stresses. For example, in mammals, p53 is activated by DNA damage (1-4) and hypoxia (5, 6), among others, and activates genes that impact cell cycle progression (7), DNA repair (8, 9), apoptosis, and angiogenesis (10, 11). In yeast, Msn2/4 responds to nutritional stress (12), as well as to salt (13), calcium (14), heat, and other stresses (15). Bacteria also contain general stress response pathways, including the alternative sigma factors RpoS in Escherichia coli (16) and σ B in Bacillus subtilis (17).It has been proposed that general stress response pathways enable cells to cross-protect, by anticipating stresses that may not be present at the moment, but are likely to occur soon (18). For example, preexposure to specific stresses is known to enhance bacterial resistance to different stresses applied subsequently (19)(20)(21). This raises a basic question: How do cells determine when to use the general stress response rather than activating more specific individual pathways?The σ B -mediated general stress response of B. subtilis provides an ideal model system to address these issues. σ B is activated by diverse stresses through a well-characterized and conserved transcriptional and posttranscriptional circuit mechanism (17). In response to stress, it activates ∼200 target ge...

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“…These systems are sufficient to provide osmoprotection to growing cells (9). However, the osmospecific stress response is also linked to the general stress response because at least one transport system for compatible solutes (opuE) is also subject to the control of B (51,60). While osmospecific protection systems provide protection to growing cells, the general stress regulon's protective role seems to be predominantly confined to nongrowing cells (59).…”

Section: Vol 187 2005 General Stress Genes and Stress Resistance 2823mentioning

confidence: 99%

“…The proteins that have been studied include GtaB, a UDP-glucose pyrophosphohydrolase involved in cell wall synthesis (57); the ferritin-like Dps protein, which is essential for the development of stationaryphase induced resistance to oxidative stress (7); OpuE, which is required for uptake of the osmoprotectant proline (60); the essential thioredoxin encoded by trxA (46); and ClpC, the ATPase subunit of the Clp chaperone/protease that is involved in development of resistance against high temperature and salt stress (32,39). In the last three cases gene expression is not solely dependent on B but is subject to additional control mechanisms (15,31,46,51,60). However, the B regulon also comprises more than 100 genes with undefined functions that are probably also involved in the development of nonspecific and multiple-stress resistance.…”

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confidence: 99%

Comprehensive Characterization of the Contribution of Individual SigB-Dependent General Stress Genes to Stress Resistance ofBacillus subtilis

Höper¹,

Völker

Hecker³

2005

J Bacteriol

114

129

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TheB -dependent general stress regulon of Bacillus subtilis comprises more than 150 members. Induction of this regulon by imposition of environmental or metabolic stress confers multiple, nonspecific, and preemptive stress resistance to nongrowing, nonsporulated cells of B. subtilis. In this study we performed a regulonwide phenotypic screening analysis to determine the stress sensitivity profiles of 94 mutants defective in candidate members of the general stress regulon that were previously identified in our transcriptional profiling study of the general stress response of B. subtilis. The phenotypic screening analysis included analysis of adaptation to a growth-inhibiting concentration of ethanol (10%, vol/vol) or NaCl (10%, wt/vol), severe heat shock (54°C), and low temperature (survival at 4°C and growth at 12.5°C). Surprisingly, 85% of the mutants tested displayed increased sensitivity at an ␣ confidence level of <0.01 to at least one of the four stresses tested, and 62% still exhibited increased sensitivity at an ␣ of <0.001. In essence, we were able to assign 63 genes (28 genes with an ␣ of <0.001) to survival after ethanol shock, 37 genes (28 genes with an ␣ of <0.001) to protection from NaCl shock, 34 genes (24 genes with an ␣ of <0.001) to survival at 4°C, and 10 genes (3 genes with an ␣ of <0.001) to management of severe heat shock. Interestingly, there was a substantial overlap between the genes necessary for survival during ethanol shock and the genes necessary for survival at 4°C, and there was also an overlap between genes required for survival during ethanol shock and genes required for survival during NaCl shock. Our data provide evidence for the importance of the B regulon at low temperatures, not only for growth but also for survival. Moreover, the data imply that a secondary oxidative stress seems to be a common component of the severe stresses tested.

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Osmoregulation of the opuE proline transport gene from Bacillus subtilis: contributions of the sigma A‐ and sigma B‐dependent stress‐responsive promoters

Cited by 71 publications

References 53 publications

T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis

T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis

Rate of environmental change determines stress response specificity

Comprehensive Characterization of the Contribution of Individual SigB-Dependent General Stress Genes to Stress Resistance ofBacillus subtilis

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