The ftsH gene of Bacillus subtilis is involved in major cellular processes such as sporulation, stress adaptation and secretion.
The ftsH gene of Bacillus subtilis has been identified as a general stress gene which is transiently induced after thermal or osmotic upshift. The FtsH protein exhibits 70.1% homology to FtsH of Escherichia coli which constitutes an essential ATP- and Zn(2+)-dependent protease anchored in the cytoplasmic membrane via two N-terminal transmembrane domains. This paper describes the isolation and functional characterization of an ftsH null mutant which was obtained by integration of a cat-cassette near the 5' end of ftsH, thereby preventing the synthesis of FtsH protein. In contrast to the situation in E. coli, ftsH is dispensable in B. subtilis but results in a pleiotropic phenotype. While the mutant cells grew mostly as large filaments under physiological conditions, they turned out to be extremely sensitive to heat and salt stress. Although ftsH is necessary for adaptation to heat, it is not involved in the regulation of the heat-shock response. The induction profiles of representative genes of the CIRCE and sigma-B regulon and class III heat-shock genes ion and clpC were identical in the wild type and the ftsH null mutant. Furthermore, the ftsH knockout strain was unable to sporulate, and this failure was probably due to the absence of Spo0A protein which is essential for entry into the sporulation programme. In addition, secretion of bulk exoproteins was severely impaired in the ftsH null mutant after entry into stationary phase. The alpha-amylase and subtilisin activity in the supernatant was specifically tested. Whereas the activity of alpha-amylase increased after entry into stationary phase in both the wild type and the ftsH mutant strain, that of subtilisin encoded by aprE was prevented at the level of transcription in the mutant. Most of these results can be explained by the failure to synthesize appropriate amounts of Spo0A protein in the ftsH null mutant and point to ftsH as a developmental checkpoint.
Deuerling
E
E
Institute of Genetics, University of Bayreuth, Germany.
Mogk
A
A
Richter
C
C
Purucker
M
M
Schumann
W
W
D049069:ATP-Dependent Proteases; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry; D018834:Chaperonin 60 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics; D007763:Lac Operon; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D026901:Membrane Transport Proteins; D016254:Mutagenesis, Insertional; D009995:Osmosis / Q000502:physiology; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012965:Sodium Chloride / Q000494:pharmacology; D013171:Spores, Bacterial / Q000502:physiology; D013381:Subtilisins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D000516:alpha-Amylases / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
C088074:AprE protein, Bacteria; D001426:Bacterial Proteins; D018834:Chaperonin 60; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C080950:Spo0A protein, Bacillus subtilis; D014157:Transcription Factors; D012965:Sodium Chloride; D000516:alpha-Amylases; D001616:beta-Galactosidase; D010450:Endopeptidases; D049069:ATP-Dependent Proteases; C513383:FtsH protein, E coli; D013381:Subtilisins; C063842:dnaK protein, E coli