Expression of the major outer-membrane porins in is transcriptionally controlled during nutrient limitation. Expression of was more than 40-fold higher under glucose limitation than under nitrogen (ammonia) limitation in chemostat cultures at the same growth rate. In contrast, 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 , , and , as well as the two-component system, was studied under glucose and N limitation in chemostat cultures. A major contributor to low 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 transcription, but little change was observed with . Lack of RpoS under glucose limitation had a lesser stimulatory effect on expression. Porin production was minimally dependent on EnvZ under N limitation due to OmpR phosphorylation by acetyl phosphate. Evidence obtained with and mutants suggested that the acetyl phosphate level also regulates porins independently and indirectly via RpoS and other pathways. 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 mutant hardly expressed under glucose limitation but boosted . In addition, the role of DNA-binding proteins encoded by was tested under glucose limitation: the mutation reduced the glucose-limitation peak, but the mutation suppressed the 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 expression under glucose limitation.


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