Abstract
LuxR is required for cell density-dependent activation of the Vibrio fischeri luminescence (lux) genes. It has not been possible to study full-length LuxR in vitro, but a polypeptide containing the C-terminal transcriptional-activator domain of LuxR (LuxRdeltaN) has been purified, and its binding to lux regulatory DNA has been investigated. By itself, LuxRdeltaN interacts with a region of lux regulatory DNA that is upstream of the lux box, which is a 20-bp element that is required for LuxR activation of the luminescence operon. Individually, neither the purified LuxRdeltaN nor RNA polymerase binds to the lux box region, but together the two proteins bind in synergy to the lux box-luxI promoter region. We show that binding of LuxRdeltaN to the upstream region is not a prerequisite for its synergistic binding with RNA polymerase to the lux box and the luxI promoter region. We also show that LuxRdeltaN and RNA polymerase are both required and sufficient for transcriptional activation of the lux operon. This argues against the hypothesis that LuxR functions to alleviate repression of the lux operon by another cellular factor. Rather, our data support the view that LuxR functions as an accessory factor that enables RNA polymerase to bind to and initiate transcription from the promoter of the lux operon.
MeSH Terms
Bacterial Proteins/genetics,metabolism
DNA-Binding Proteins/genetics,metabolism
DNA-Directed RNA Polymerases/metabolism
Luminescent Measurements
Peptide Chain Initiation, Translational
Promoter Regions, Genetic
Repressor Proteins
Temperature
Trans-Activators/genetics,metabolism
Transcription, Genetic
Vibrio/genetics,metabolism
Chemicals
Bacterial Proteins
DNA-Binding Proteins
Repressor Proteins
Trans-Activators
LuxR autoinducer binding proteins
DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Stevens A M
Department of Microbiology, University of Iowa, Iowa City 52242, USA.
Greenberg E P
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