DksA affects ppGpp induction of RpoS at a translational level.
The RpoS sigma factor (also called sigmaS or sigma38) is known to regulate at least 50 genes in response to environmental sources of stress or during entry into stationary phase. Regulation of RpoS abundance and activity is complex, with many factors participating at multiple levels. One factor is the nutritional stress signal ppGpp. The absence of ppGpp blocks or delays the induction of rpoS during entry into stationary phase. Artificially inducing ppGpp, without starvation, is known to induce rpoS during the log phase 25- to 50-fold. Induction of ppGpp is found to have only minor effects on rpoS transcript abundance or on RpoS protein stability; instead, the efficiency of rpoS mRNA translation is increased by ppGpp as judged by both RpoS pulse-labeling and promoter-independent effects on lacZ fusions. DksA is found to affect RpoS abundance in a manner related to ppGpp. Deleting dksA blocks rpoS induction by ppGpp. Overproduction of DksA induces rpoS but not ppGpp. Deleting dksA neither alters regulation of ppGpp in response to amino acid starvation nor nullifies the inhibitory effects of ppGpp on stable RNA synthesis. Although this suggests that dksA is epistatic to ppGpp, inducing ppGpp does not induce DksA. A dksA deletion does display a subset of the same multiple-amino-acid requirements found for ppGpp(0) mutants, but overproducing DksA does not satisfy ppGpp(0) requirements. Sequenced spontaneous extragenic suppressors of dksA polyauxotrophy are frequently the same T563P rpoB allele that suppresses a ppGpp(0) phenotype. We propose that DksA functions downstream of ppGpp but indirectly regulates rpoS induction.
Brown
Larissa
L
Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, Bethesda, Maryland 20892, USA.
Gentry
Daniel
D
Elliott
Thomas
T
Cashel
Michael
M
D001426:Bacterial Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D029968:Escherichia coli Proteins; D017353:Gene Deletion; D018628:Gene Dosage; D015964:Gene Expression Regulation, Bacterial; D016153:Genes, Suppressor / Q000502:physiology; D006159:Guanosine Tetraphosphate / Q000172:deficiency / Q000378:metabolism*; D007763:Lac Operon / Q000235:genetics; D008025:Ligases / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis / Q000502:physiology; D010641:Phenotype; D014176:Protein Biosynthesis / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012808:Sigma Factor / Q000235:genetics*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012808:Sigma Factor; C090751:dksA protein, E coli; C069567:sigma factor KatF protein, Bacteria; D006159:Guanosine Tetraphosphate; D008025:Ligases; C028812:guanosine 3',5'-polyphosphate synthetases