Membrane protein overproduction stress in Bacillus subtilis

Comparison of the transcriptome of Bacillus subtilis under going membrane protein overproduction in the wild type, sigW and cssRS deletion strains. We demonstrate that the dynamics of the stress systems involved in membrane overproduction are far more complicated than was first hypothesised and that many more systems than SigW and CssRS are involved in membrane protein overexpression stress. Interestingly the cssRS genes are repressed in the sigW deletion strain. RNA samples prepared from B. subtilis 168 amyE::spaRK pNZ8910 (empty plasmid), 168 amyE::spaRK pNS8910::B.plsY and mutants sigW amyE::spaRK pNZ8910::B.plsY and cssRS amyE spaRK pNZ8910::B.plsY were grown in Luria Bertani broth to O.D 600nm 1.0 before being induced with 1% subtilin and grown for a futher 2 hours were reverse transcribed, labeled and hybridized to tiling arrays. All hybridizations were performed in triplicate using RNA isolated from independent cultures.