Control of Vibrio fischeri lux gene transcription by a cyclic AMP receptor protein-luxR protein regulatory circuit.

AUTOR(ES)

Dunlap, P V

RESUMO

Expression of the Vibrio fischeri luminescence genes (lux genes) requires two transcriptional activators: the V. fischeri luxR gene product with autoinducer and the cyclic AMP (cAMP) receptor protein (CRP) with cAMP. It has been established that autoinducer and the luxR gene product are required for transcriptional activation of the luxICDABE operon, which contains a gene required for autoinducer synthesis and genes required for light emission. However, the role of cAMP-CRP in the induction of luminescence is not clear. We examined transcriptional control of the lux genes in Escherichia coli, using catabolite repression mutants carrying lux DNA-containing plasmids. Transcriptional fusions between the lacZ gene on Mu dI and luxR were used to assess luxR promoter activity, and the luxAB genes (which encode the two luciferase subunits) were used as a natural reporter of luxICDABE promoter activity. A plasmid containing luxR under control of the cAMP-CRP-independent tac promoter was constructed to direct the synthesis of the luxR gene product in cells containing compatible luxR::Mu dI insertion mutant plasmids. In E. coli, cAMP-CRP activated transcription of luxR and concurrently decreased luxICDABE transcription. In the presence of relatively high levels of the luxR gene product, cAMP and CRP were not required for induction of the luxICDABE operon. The luxR gene product in the presence of autoinducer activated transcription of the luxICDABE operon, as has been shown previously, and we demonstrate that it also decreased luxR transcription. Apparently, control of the V. fischeri luminescence genes involves a regulatory circuit in which cAMP and CRP activate luxR transcription and in turn the luxR gene product activates transcription of the operon responsible for light emission (uxICDABE). Furthermore, in lux gene regulation cAMP-CRP and autoinducer-LuxR protein appear to function as transcriptional antagonists.

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