An analysis of multifactorial influences on the transcriptional control of ompF and ompC porin expression under nutrient limitation.
Expression of the major outer-membrane porins in Escherichia coli is transcriptionally controlled during nutrient limitation. Expression of ompF was more than 40-fold higher under glucose limitation than under nitrogen (ammonia) limitation in chemostat cultures at the same growth rate. In contrast, ompC expression was higher under N limitation. The basis of regulation by nutrient limitation was investigated using mutations affecting expression of porin genes. The influence of cyaA, rpoS, ackA and pta, as well as the two-component envZ-ompR system, was studied under glucose and N limitation in chemostat cultures. A major contributor to low ompF expression under N limitation was negative control by the RpoS sigma factor. RpoS levels were high under N limitation and loss of RpoS resulted in a 19-fold increase in ompF transcription, but little change was observed with ompC. Lack of RpoS under glucose limitation had a lesser stimulatory effect on ompF expression. Porin production was minimally dependent on EnvZ under N limitation due to OmpR phosphorylation by acetyl phosphate. Evidence obtained with pta and ackA mutants suggested that the acetyl phosphate level also regulates porins independently and indirectly via RpoS and other pathways. pta-envZ double mutants had a residual level of porin transcription, implicating alternative means of OmpR phosphorylation under nutrient limitation. Another critical factor in regulation was the level of cAMP, as a cyaA mutant hardly expressed ompF under glucose limitation but boosted ompC. In addition, the role of DNA-binding proteins encoded by hns and himA was tested under glucose limitation: the hns mutation reduced the glucose-limitation peak, but the himA mutation suppressed the hns effect, suggesting a complex web of interrelationships between the DNA-binding proteins. Indeed, multiple inputs and no single regulator were responsible for the high peak of ompF expression under glucose limitation.
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Department of Microbiology G08, University of Sydney, Sydney, New South Wales 2006, Australia.
Ferenci
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D000262:Adenylyl Cyclases / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics*; D002463:Cell Membrane Permeability; D003470:Culture Media; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D009154:Mutation; D010755:Organophosphates / Q000378:metabolism; D010709:Phosphate Acetyltransferase / Q000235:genetics / Q000378:metabolism; D018272:Porins / Q000235:genetics*; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D003470:Culture Media; D004268:DNA-Binding Proteins; C031908:H-NS protein, bacteria; C113724:OmpC protein; C109237:OmpF protein; D010755:Organophosphates; D018272:Porins; D011993:Recombinant Fusion Proteins; C011632:acetyl phosphate; D010709:Phosphate Acetyltransferase; D000262:Adenylyl Cyclases; D005947:Glucose