Bacillus subtilis W168, WT vs. sigA (G336C)
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ABSTRACT:
ORGANISM(S): Bacillus subtilis
SUBMITTER: John Helmann
PROVIDER: E-GEOD-55202 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Transcriptional profile of the uppS-RBS (A to C) mutant during log growth phase in LB medium. Bacillus subtilis W168, WT vs. uppS-RBS (A to C). The experiment was conducted two times using three independent total RNA preparations (biological triplicates). Each comparison was performed in dye-swap with the same RNA preparation. For each paried comparison, one sample was labeled with Alexa Fluor 555 and the other was with Alexa Fluor 647.
2013-04-01 | E-GEOD-45305 | biostudies-arrayexpress
Project description:Transcriptional profile of the rocG gudB double null mutant during log growth phase in LB medium. Bacillus subtilis W168, WT vs. delta-rocG delta-gudB. The experiment was conducted two times using three independent total RNA preparations (biological triplicates). For each paried comparison, WT was labeled with Alexa Fluor 555 and the rocG gudB double mutant was with Alexa Fluor 647.
2011-12-13 | E-GEOD-34383 | biostudies-arrayexpress
Project description:Transcriptional response of Bacillus subtilis to daptomycin in wild-type and in a daptomycin resistant mutant. Bacillus subtilis 168, WT (-DAP) vs. DapR1 (-DAP), WT (+DAP) vs. DapR1 (+DAP), DapR1 (+DAP) vs. DapR1 (-DAP). Each experiment was conducted at least twice using two independent total RNA preparations. For daptomycin untreated comparison between 168 WT and DapR1 mutant, DapR1 was labeled with Alexa Fluor 647 and WT was labeled with Alexa Fluor 555. For daptomycin treated experiments between WT and DapR1, DapR1 was labeled with Alexa Fluor 647 and WT with Alexa Fluor 555. For treated vs. untreated DapR1, the DAP treated samples were labeled with Alexa Fluor 647 and the untreated with Alexa Fluor 555. For dye swap, untreated DapR1 was labeled with Alexa Fluor 647 and DAP treated with Alexa Fluor 555.
2011-06-16 | E-GEOD-30000 | biostudies-arrayexpress
Project description:The Bacillus subtilis membrane contains diacylglycerol-based lipids with at least five distinct headgroups that impart distinct physical and chemical properties to the lipid bilayer. Here, we describe the phenotypic characterization of mutant strains lacking one or more of the following lipids: glycolipids (ugtP mutants), phosphatidylethanolamine (pssA and psd mutants), lysylphosphatidylglycerol (mprF), and cardiolipin (ywnE and ywjE). Alterations of membrane lipid headgroup composition are generally well tolerated by the cell and even severe alterations lead to only modest effects on growth proficiency. Mutants with decreased levels of positively charged lipids display an increased sensitivity to a cationic antimicrobial compounds and cells lacking glycolipids are sensitive to the lantibiotic sublancin and are defective in swarming motility. A quadruple mutant strain (ugtP pssA mprF ywnE), with a membrane comprised predominantly of phosphatidylglycerol, is viable and grows at near wild-type rates, although it forms long, coiled filaments. Transcriptome comparisons identify numerous regulons with altered expression in ugtP mutant cells, the pssA mprF ywnE triple mutant, and the uptP pssA mprF ywnE quadruple mutant. These effects include a general decrease in expression of the SigD and FapR regulons, and increased expression of cell envelope stress responses mediated by ?M and the YvrGHb two-component system. WT vs.ugtP single mutant (U), WT vs. mprFpssAywnE triple mutant (T) or WT vs. ugtPmprFpssAywnE quadruple mutant (Q). Each experiment (WT vs. mutant) was conducted four times using three independent total RNA preparations (3 independent experiement + dye swap). For the 3 datasets used for final analysis wild-type was labeled with Alexa Fluor 555 and mutants with Alexa Fluor 647. For dye swap experiments wild-type was labeled with Alexa Fluor 647 and mutants with Alexa Fluor 555.
2010-05-18 | E-GEOD-13036 | biostudies-arrayexpress
Project description:Abstract of associated manuscript: The Bacillus subtilis extracytoplasmic function (ECF) sigma(M) factor is activated by cell envelope stress elicited by antibiotics, and by acid, heat, ethanol and superoxide stresses. Here, we have used several complementary approaches to identify genes controlled by sigma(M). In many cases, expression is only partially dependent on sigma(M) because of both overlapping promoter recognition with other ECF sigma factors and the presence of additional promoter elements. Genes regulated by sigma(M) have a characteristic pattern of induction in response to cell envelope-acting antibiotics as evidenced by hierarchical clustering analysis. sigma(M) also contributes to the expression of the Spx transcription factor and thereby indirectly regulates genes of the Spx regulon. Cell envelope stress responses also include regulons controlled by sigma(W), sigma(B) and several two-component regulatory systems (e.g. LiaRS, YycFG, BceRS). Activation of the sigma(M) regulon increases expression of proteins functioning in transcriptional control, cell wall synthesis and shape determination, cell division, DNA damage monitoring, recombinational repair and detoxification. WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van). Each experiment was conducted at least twice using two independent total RNA preparations. For vancomycin untreated and treated experiments, untreated samples were labeled with Alexa Fluor 555 and treated samples with Alexa Fluor 647. For WT vs. mutant experiments, wild type was labeled with Alexa Fluor 555 and mutants with Alexa Fluor 647. For dye swap experiment, wild-type was labeled with Alexa Fluor 647 and mutant with Alexa Fluor 555. Bacillus subtilis CU1065, WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van)
2010-05-15 | E-GEOD-13820 | biostudies-arrayexpress
Project description:Bacillus subtilis encodes seven extracytoplasmic function (ECF) sigma factors. Three (sigma M, sigma W and simga X) mediate responses to cell envelope active antibiotics. The functions of sigma Y, sigma Z, sigma V, and YlaC remain largely unknown, and strong inducers of these sigma factors and their regulons have yet to be defined. Here, we define transcriptomic and phenotypic differences under non-stress conditions between strains carrying deletions in all seven ECF sigma factor genes (Δ7ECF), a sigMWX triple mutant (∆MWX), and the parental 168 strain. Our results identify >80 genes as at least partially dependent on ECF sigma factors and, as expected, most of these are dependent on sigma M, sigma W or sigma X which are active at a significant basal level during growth. Several genes, including the eps operon encoding enzymes for exopolysaccharide (EPS) production, were decreased in expression in Δ7ECF but affected little if at all in ΔMWX. Consistent with this observation, Δ7ECF (but not ∆MWX) showed reduced biofilm formation. Extending previous observations, we also note that ∆MWX is sensitive to a variety of antibiotics and Δ7ECF is either as sensitive as, or slightly more sensitive than, the ΔMWX strain to these stressors. These findings emphasize the overlapping nature of the seven ECF s factor regulons in B. subtilis, confirm that three of these (sigma M, W or X) play the dominant role in conferring intrinsic resistance to antibiotics, and provide initial insights into the roles of the remaining ECF sigma factors. Strains WT vs. ΔMWX, WT vs. Δ7ECF, Δ7ECF vs. ΔMWX. Each experiment was conducted three times using three independent total RNA preparations (biological triplicates). For each paried comparison, one sample was was labeled with Alexa Fluor 555 and the other was with Alexa Fluor 647. For each comparison, one replicate was performed with dyewap with the same RNA preparation.
2010-07-21 | E-GEOD-22930 | biostudies-arrayexpress
Project description:Transcriptional response of Bacillus subtilis to moenomycin in wild-type 168. Bacillus subtilis 168, WT (-MOE) vs. WT (+MOE). The experiment was conducted in triplicate using three independent total RNA preparations. Untreated samples were labeled with Alexa Fluor 555 and moenomycin treated samples were labeled with Alexa Fluor 647.
2011-06-17 | E-GEOD-30001 | biostudies-arrayexpress
Project description:Abstract of associated manuscript: Daptomycin is the first of a new class of cyclic lipopeptide antibiotics used against multidrug-resistant Gram-positive pathogens. The proposed mechanism of action involves disruption of the functional integrity of the bacterial membrane in a Ca2+-dependent manner. We have used transcriptional profiling to demonstrate that treatment of Bacillus subtilis with daptomycin strongly induces the lia operon including the autoregulatory LiaRS two-component system (homologous to Staphylococcus aureus VraSR). The lia operon protects against daptomycin and deletion of liaH, encoding a phage shock protein A (PspA)-like protein, leads to 3-fold increased susceptibility. Since daptomycin interacts with the membrane, we tested mutants with altered membrane composition for effects on susceptibility. Deletion mutations of mprF (lacking lysyl-phosphatidylglycerol) or des (lipid desaturase) increased daptomycin susceptibility, whereas overexpression of MprF decreased susceptibility. Conversely, depletion of the cell for the anionic lipid phosphatidylglycerol led to increased resistance. Fluorescently-labeled daptomycin localized to the septa and in a helical pattern around the cell envelope and was delocalized upon depletion of phosphatidylglycerol. Together, these results indicate that the daptomycin-Ca2+ complex interacts preferentially with regions enriched in anionic phospholipids and leads to membrane stresses that can be ameliorated by PspA family proteins. Bacillus subtilis W168, WT (+DAP) vs. WT (-DAP). The experiment was conducted in triplicate using three independent total RNA preparations. For WT-rep1 and WT-rep2, daptomycin treated samples were labeled with Alexa Fluor 647 and untreated samples with Alexa Fluor 555. For WT-rep3, the daptomycin treated sample was labeled with Alexa Fluor 555 and the untreated sample with Alexa Fluor 647.
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