scholarly journals An Stomatin, Prohibitin, Flotillin, and HflK/C-Domain Protein Required to Link the Phage-Shock Protein to the Membrane in Bacillus subtilis

2021 ◽  
Vol 12 ◽  
Author(s):  
Abigail Savietto Scholz ◽  
Sarah S. M. Baur ◽  
Diana Wolf ◽  
Marc Bramkamp
Keyword(s):  
Bacillus Subtilis ◽  
Membrane Fluidity ◽  
Cell Envelope ◽  
Cellular Life ◽  
Fluid Membrane ◽  
Envelope Stress ◽  
Spfh Domain ◽  
Domain Protein ◽  
Integral Proteins ◽  
Detergent Resistant Membrane

Membrane surveillance and repair is of utmost importance to maintain cellular integrity and allow cellular life. Several systems detect cell envelope stress caused by antimicrobial compounds and abiotic stresses such as solvents, pH-changes and temperature in bacteria. Proteins containing an Stomatin, Prohibitin, Flotillin, and HflK/C (SPFH)-domain, including bacterial flotillins have been shown to be involved in membrane protection and membrane fluidity regulation. Here, we characterize a bacterial SPFH-domain protein, YdjI that is part of a stress induced complex in Bacillus subtilis. We show that YdjI is required to localize the ESCRT-III homolog PspA to the membrane with the help of two membrane integral proteins, YdjG/H. In contrast to classical flotillins, YdjI resides in fluid membrane regions and does not enrich in detergent resistant membrane fractions. However, similarly to FloA and FloT from B. subtilis, deletion of YdjI decreases membrane fluidity. Our data reveal a hardwired connection between phage shock response and SPFH proteins.

scholarly journals The Cell Envelope Stress Response of Bacillus subtilis towards Laspartomycin C

Antibiotics ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 729
Author(s):  
Angelika Diehl ◽  
Thomas M. Wood ◽  
Susanne Gebhard ◽  
Nathaniel I. Martin ◽  
Georg Fritz
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Stress Response ◽  
Cell Envelope ◽  
Resistance Mechanisms ◽  
Detailed Knowledge ◽  
Knock Out ◽  
Lipid Ii ◽  
Envelope Stress ◽  
The Impact

Cell wall antibiotics are important tools in our fight against Gram-positive pathogens, but many strains become increasingly resistant against existing drugs. Laspartomycin C is a novel antibiotic that targets undecaprenyl phosphate (UP), a key intermediate in the lipid II cycle of cell wall biosynthesis. While laspartomycin C has been thoroughly examined biochemically, detailed knowledge about potential resistance mechanisms in bacteria is lacking. Here, we use reporter strains to monitor the activity of central resistance modules in the Bacillus subtilis cell envelope stress response network during laspartomycin C attack and determine the impact on the resistance of these modules using knock-out strains. In contrast to the closely related UP-binding antibiotic friulimicin B, which only activates ECF σ factor-controlled stress response modules, we find that laspartomycin C additionally triggers activation of stress response systems reacting to membrane perturbation and blockage of other lipid II cycle intermediates. Interestingly, none of the studied resistance genes conferred any kind of protection against laspartomycin C. While this appears promising for therapeutic use of laspartomycin C, it raises concerns that existing cell envelope stress response networks may already be poised for spontaneous development of resistance during prolonged or repeated exposure to this new antibiotic.

scholarly journals Daptomycin versus Friulimicin B: In-Depth Profiling of Bacillus subtilis Cell Envelope Stress Responses

2009 ◽  
Vol 53 (4) ◽  
pp. 1619-1623 ◽  
Author(s):  
Tina Wecke ◽  
Daniela Zühlke ◽  
Ulrike Mäder ◽  
Sina Jordan ◽  
Birgit Voigt ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Stress Responses ◽  
Cell Envelope ◽  
Depth Profiling ◽  
Peptide Antibiotics ◽  
Two Component System ◽  
Genome Wide ◽  
Envelope Stress ◽  
Σ Factor ◽  
Stress Sensing

ABSTRACT The related lipo(depsi)peptide antibiotics daptomycin and friulimicin B show great potential in the treatment of multiply resistant gram-positive pathogens. Applying genome-wide in-depth expression profiling, we compared the respective stress responses of Bacillus subtilis. Both antibiotics target envelope integrity, based on the strong induction of extracytoplasmic function σ factor-dependent gene expression. The cell envelope stress-sensing two-component system LiaRS is exclusively and strongly induced by daptomycin, indicative of different mechanisms of action in the two compounds.

scholarly journals Cell Envelope Stress Response in Cell Wall-Deficient L-Forms of Bacillus subtilis

2012 ◽  
Vol 56 (11) ◽  
pp. 5907-5915 ◽  
Author(s):  
Diana Wolf ◽  
Patricia Domínguez-Cuevas ◽  
Richard A. Daniel ◽  
Thorsten Mascher
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cell Envelope ◽  
Membrane Integrity ◽  
Culture Conditions ◽  
Content Type ◽  
Damage Sensing ◽  
Controlled Switching ◽  
Gene Assay ◽  
Envelope Stress

ABSTRACTL-forms are cell wall-deficient bacteria that can grow and proliferate in osmotically stabilizing media. Recently, a strain of the Gram-positive model bacteriumBacillus subtiliswas constructed that allowed controlled switching between rod-shaped wild-type cells and corresponding L-forms. Both states can be stably maintained under suitable culture conditions. Because of the absence of a cell wall, L-forms are known to be insensitive to β-lactam antibiotics, but reports on the susceptibility of L-forms to other antibiotics that interfere with membrane-anchored steps of cell wall biosynthesis are sparse, conflicting, and strongly influenced by strain background and method of L-form generation. Here we investigated the response ofB. subtilisto the presence of cell envelope antibiotics, with regard to both antibiotic resistance and the induction of the known LiaRS- and BceRS-dependent cell envelope stress biosensors. Our results show thatB. subtilisL-forms are resistant to antibiotics that interfere with the bactoprenol cycle, such as bacitracin, vancomycin, and mersacidin, but are hypersensitive to nisin and daptomycin, which both affect membrane integrity. Moreover, we established alacZ-based reporter gene assay for L-forms and provide evidence that LiaRS senses its inducers indirectly (damage sensing), while the Bce module detects its inducers directly (drug sensing).

scholarly journals Regulation of LiaRS-Dependent Gene Expression in Bacillus subtilis: Identification of Inhibitor Proteins, Regulator Binding Sites, and Target Genes of a Conserved Cell Envelope Stress-Sensing Two-Component System

2006 ◽  
Vol 188 (14) ◽  
pp. 5153-5166 ◽  
Author(s):  
Sina Jordan ◽  
Anja Junker ◽  
John D. Helmann ◽  
Thorsten Mascher
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Target Genes ◽  
Cell Envelope ◽  
Protein A ◽  
Negative Regulator ◽  
Genomic Context ◽  
Envelope Stress ◽  
Two Component ◽  
Stress Signals

ABSTRACT The regulatory network of the cell envelope stress response in Bacillus subtilis involves both extracytoplasmic function σ-factors and two-component signal transducing systems. One such system, LiaRS, responds to cell wall antibiotics that interfere with the undecaprenol cycle and to perturbation of the cytoplasmic membrane. It is encoded by the last two genes of the liaIHGFSR locus. Here, we analyzed the expression of two LiaR-dependent operons, liaIHGFSR and yhcYZ-yhdA, and characterized a palindromic sequence required for LiaR-dependent activation. Since induction of the strong liaI promoter leads to both liaIH and liaIHGFRS transcripts, LiaR is positively autoregulated. Systematic deletion analysis of the liaI operon revealed that LiaF is a potent negative regulator of LiaR-dependent gene expression: a nonpolar liaF deletion led to constitutive activation of both characterized LiaR-dependent promoters. The liaF gene is conserved in both sequence and genomic context in the Firmicutes group of gram-positive bacteria, located directly upstream of liaSR orthologs. LiaH, a homolog of Escherichia coli phage shock protein A, also plays a more subtle role in negatively modulating the bacitracin-inducible expression from LiaR-dependent promoters. Our results support a model in which the LiaFRS module integrates both positive and negative feedback loops to transduce cell envelope stress signals.

scholarly journals Extracytoplasmic Function σ Factors with Overlapping Promoter Specificity Regulate Sublancin Production in Bacillus subtilis

2009 ◽  
Vol 191 (15) ◽  
pp. 4951-4958 ◽  
Author(s):  
Yun Luo ◽  
John D. Helmann
Keyword(s):  
Bacillus Subtilis ◽  
Transcriptional Repression ◽  
Cell Envelope ◽  
Antibiotic Production ◽  
Ectopic Expression ◽  
Genetic Studies ◽  
Envelope Stress ◽  
Positive Regulators ◽  
State Regulator ◽  
Transition State Regulator

ABSTRACT Bacillus subtilis harbors seven extracytoplasmic function (ECF) σ factors. At least three ECF σ factors (σM, σW, and σX) are induced by, and provide resistance to, antibiotics and other agents eliciting cell envelope stress. Here, we report that ECF σ factors also contribute to antibiotic production. B. subtilis 168 strains that are lysogenic for the SPβ bacteriophage produce sublancin, which inhibits the growth of other, nonlysogenic strains. Genetic studies demonstrate that synthesis of sublancin is largely dependent on σX, with a smaller contribution from σM. A sigM sigX double mutant is unable to produce sublancin. This defect is primarily due to the fact that the sublancin biosynthesis is positively activated by the transition state regulator and AbrB paralog Abh, which counteracts transcriptional repression of the sublancin biosynthesis operon by AbrB. Ectopic expression of abh bypasses the requirement for σM or σX in sublancin synthesis, as does an abrB mutation. In addition to their major role in regulating sublancin expression by activating abh transcription, σX and σM also have a second role as positive regulators of sublancin expression that is independent of AbrB and Abh. Since sublancin resistance in nonlysogens is largely dependent on σW, ECF σ factors control both sublancin production and resistance.

scholarly journals An Antibiotic-Inducible Cell Wall-Associated Protein That Protects Bacillus subtilis from Autolysis

2007 ◽  
Vol 189 (13) ◽  
pp. 4671-4680 ◽  
Author(s):  
Letal I. Salzberg ◽  
John D. Helmann
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cell Envelope ◽  
Sequence Similarity ◽  
Envelope Stress ◽  
Class B ◽  
Dependent Cell ◽  
A Cell ◽  
Close Relatives ◽  
Minimal Region

ABSTRACT In Bacillus subtilis, antibiotics that impair cell wall synthesis induce a characteristic stress response including the σW and σM regulons and the previously uncharacterized yoeB gene. Here we demonstrate that YoeB is a cell wall-associated protein with weak sequence similarity to a noncatalytic domain of class B penicillin-binding proteins. A yoeB-null mutant exhibits an increased rate of autolysis in response to cell wall-targeting antibiotics or nutrient depletion. This phenotype does not appear to be correlated with gross alterations in peptidoglycan structure or levels of autolysins. Promoter dissection experiments define a minimal region necessary for antibiotic-mediated induction of yoeB, and this region is highly conserved preceding yoeB homologs in close relatives of B. subtilis. These results support a model in which induction of YoeB in response to cell envelope stress decreases the activity of autolysins and thereby reduces the rate of antibiotic-dependent cell death.

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