The ftsH gene of Bacillus subtilis is transiently induced after osmotic and temperature upshift

Abstract

The ftsH gene of Bacillus subtilis has been identified as a salt-sensitive insertion mutation in strain UG1. Here, we show that UG1 has an insertion near the 3' end of ftsH. The salt sensitivity of this mutant was caused by reduction of ftsH mRNA levels by the synthesis of an artificial antisense RNA originating at a promoter located within the insertion and reading backwards into the ftsH gene. The salt-sensitive phenotype could be overcome by deleting the promoter from which the antisense RNA was transcribed. A physiological analysis of the isogenic wild-type strain in minimal medium revealed unimpaired growth at up to 1 M NaCl, and growth above 1.2 M NaCl was observed only after addition of the osmoprotectant proline or glycine betaine. In contrast, growth of strain UG1 was reduced at a salt concentration above 0.2 M, which could be rescued by the two compatible solutes already mentioned and also by trehalose. Primer extension revealed one potential transcription start site downstream of a putative vegetative promoter, which was activated after osmotic or temperature upshift. Northern (RNA blot) experiments led to the detection of a 2.1-kb transcript, suggesting that ftsH is monocistronic. A transcriptional fusion between ftsH and the gus reporter gene exhibited a twofold increase in beta-glucuronidase activity after osmotic upshift. To further confirm the need for an enhanced level of FtsH protein after osmotic upshift, the promoter was replaced by the sucrose-inducible promoter PsacB. Whereas this mutant strain could grow in the absence of inducer in LB medium, it stopped growth immediately after addition of 1.1 M NaCl. We conclude that an increased amount of FtsH protein is essential for B. subtilis to cope with an increase in osmolarity or temperature.