Mutations that increase expression of the rpoS gene and decrease its dependence on hfq function in Salmonella typhimurium.
The RpoS transcription factor (also called sigmaS or sigma38) is required for the expression of a number of stationary-phase and osmotically inducible genes in enteric bacteria. RpoS is also a virulence factor for several pathogenic species, including Salmonella typhimurium. The activity of RpoS is regulated in response to many different signals, at the levels of both synthesis and proteolysis. Previous work with rpoS-lac protein fusions has suggested that translation of rpoS requires hfq function. The product of the hfq gene, host factor I (HF-I), is a ribosome-associated, site-specific RNA-binding protein originally characterized for its role in replication of the RNA bacteriophage Qbeta of Escherichia coli. In this study, the role of HF-I was explored by isolating suppressor mutations that map to the region directly upstream of rpoS. These mutations increase rpoS-lac expression in the absence of HF-I and also confer substantial independence from HF-I. DNA sequence analysis of the mutants suggests a model in which the RNA secondary structure near the ribosome binding site of the rpoS mRNA plays an important role in limiting expression in the wild type. Genetic tests of the model confirm its predictions, at least in part. It seems likely that the mutations analyzed here activate a suppression pathway that bypasses the normal HF-I-dependent route of rpoS expression; however, it is also possible that some of them identify a sequence element with an inhibitory function that is directly counteracted by HF-I.
Brown
L
L
Department of Microbiology and Immunology, West Virginia University Health Sciences Center, Morgantown 26506, USA.
Elliott
T
T
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D017930:Genes, Reporter; D035001:Host Factor 1 Protein; D034982:Integration Host Factors; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial; D012333:RNA, Messenger; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012808:Sigma Factor / Q000096:biosynthesis*; D013489:Suppression, Genetic
D001426:Bacterial Proteins; D002352:Carrier Proteins; D035001:Host Factor 1 Protein; D034982:Integration Host Factors; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012808:Sigma Factor; C069567:sigma factor KatF protein, Bacteria