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Global profiling of B. subtilis comparing wild-type to mutants with altered cytoplasmic membrane composition

ABSTRACT: 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.

ORGANISM(S): Bacillus subtilis

PROVIDER: GSE13036 | GEO | 2008/10/09

SECONDARY ACCESSION(S): PRJNA109709

REPOSITORIES: GEO

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Global profiling of B. subtilis comparing wild-type to mutants with altered cytoplasmic membrane composition

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

Daptomycin stimulon in Bacillus subtilis W168

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.

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Daptomycin stimulon in Bacillus subtilis W168

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Defective pgsA contributes to increased membrane fluidity and cell wall thickening in S. aureus with high-level daptomycin resistance

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2020-09-20 | GSE152567 | GEO

Bisulfite (methylome) sequencing of progenies of Col-0 grafted on msh1; Col-0 grafted on dcl2,3,4,msh1; Col-0 grafted on Col-0; msh1 mutant; dcl2,3,4,msh1 quadruple mutant in Arabidopsis [msh1_graft_BS-seq]

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2020-09-20 | GSE152564 | GEO

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