Role of anionic phospholipids in the adaptation of Bacillus subtilis to high salinity

• Autores: López, C.S.; Alice, A.F.; Heras, H.; Rivas, E.A.; Sánchez-Rivas, C.
• Año de publicación: 2006
• Idioma: inglés
• Tipo de recurso: artículo
• Fuente: Microbiology 2006;152(3):605-616
• Materia: cardiolipin; fatty acid; membrane lipid; phosphatidylglycerol; phospholipid; sodium chloride; synthetase; animal cell; article; Bacillus subtilis; bacterial growth; bacterial membrane; bacterium culture; cell vacuole; concentration (parameters); controlled study; correlation analysis; culture medium; fluorescence spectroscopy; gene insertion; gene mutation; genetic transcription; growth curve; lipid composition; lipogenesis; membrane structure; nonhuman; optical density; osmosis; priority journal; salinity; salt tolerance; steady state; strain difference; temperature; wild type; Adaptation, Physiological; Bacillus subtilis; Bacterial Proteins; Cardiolipins; Cell Membrane; Fatty Acids; Heat-Shock Response; Mutation; Osmolar Concentration; Osmotic Pressure; Phosphatidylglycerols; Sodium Chloride; Spores, Bacterial; Animalia; Bacillus subtilis; Bacteria (microorganisms)
• Institución: Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales

The importance of the content of anionic phospholipids [cardiolipin (CL) and phosphatidylglycerol (PG)] in the osmotic adaptation and in the membrane structure of Bacillus subtilis cultures was investigated. Insertion mutations in the three putative cardiolipin synthase genes (ywiE, ywnE and ywjE) were obtained. Only the ywnE mutation resulted in a complete deficiency in cardiolipin and thus corresponds to a true clsA gene. The osmotolerance of a clsA mutant was impaired: although at NaCl concentrations lower than 1.2 M the growth curves were similar to those of its wild-type control, at 1 .5 M NaCl (LBN medium) the lag period increased and the maximal optical density reached was lower. The membrane of the clsA mutant strain showed an increased PG content, at both exponential and stationary phase, but no trace of CL in either LB or LBN medium. As well as the deficiency in CL synthesis, the clsA mutant showed other differences in lipid and fatty acids content compared to the wild-type, suggesting a cross-regulation in membrane lipid pathways, crucial for the maintenance of membrane functionality and integrity. The biophysical characteristics of membranes and large unilamellar vesicles from the wild-type and clsA mutant strains were studied by Laurdan's steady-state fluorescence spectroscopy. At physiological temperature, the clsA mutant showed a decreased lateral lipid packing in the protein-free vesicles and isolated membranes compared with the wild-type strain. Interestingly, the lateral lipid packing of the membranes of both the wild-type and clsA mutant strains increased when they were grown in LBN. In a conditional IPTG-controlled pgsA mutant, unable to synthesize PG and CL in the absence of IPTG, the osmoresistance of the cultures correlated with their content of anionic phospholipids. The transcriptional activity of the clsA and pgsA genes was similar and increased twofold upon entry to stationary phase or under osmotic upshift. Overall, these results support the involvement of the anionic phospholipids in the growth of B. subtilis in media containing elevated NaCl concentrations. © 2006 SGM.
Fil:López, C.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Alice, A.F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.