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PMID: 7809088 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Synergistic binding of the Vibrio fischeri LuxR transcriptional activator domain and RNA polymerase to the lux promoter region.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 26 ·1994-12-20 ·Pages 12619-23

Stevens AM, Dolan KM, Greenberg EP

Abstract

LuxR, the Vibrio fischeri luminescence gene (lux) activator, is the best-studied member of a family of bacterial transcription factors required for cell density-dependent expression of specific genes involved in associations with eukaryotic hosts. Neither LuxR nor any other LuxR homolog has been shown to bind DNA directly. We have purified the LuxR C-terminal transcriptional activator domain from extracts of recombinant Escherichia coli in which this polypeptide was expressed. The purified polypeptide by itself binds to lux regulatory DNA upstream of the lux box, a 20-bp palindrome that is required for LuxR activity in vivo, but it does not bind to the lux box. However, the LuxR C-terminal domain together with RNA polymerase protects a region including the lux box and the lux operon promoter from DNase I cleavage. There is very little protection of the lux operon promoter region from DNase I digestion in the presence of RNA polymerase alone. Apparently, there is a synergistic binding of the LuxR C-terminal domain and RNA polymerase to the promoter region. The upstream binding region for the purified polypeptide encompasses a binding site for cAMP receptor protein (CRP). Under some conditions, CRP binding can block the binding of the LuxR C-terminal domain to the upstream binding region, and it can also block the synergistic binding of the LuxR C-terminal domain and RNA polymerase to the lux box and luminescence gene promoter region. This description of DNA binding by the LuxR C-terminal domain should lead to an understanding of the molecular interactions of the LuxR family of transcriptional activators with regulatory DNA.

Related Genes
lux luxI luxR
MeSH Terms
Bacterial Proteins/metabolism DNA-Binding Proteins/metabolism DNA-Directed RNA Polymerases/metabolism Gene Expression Regulation, Bacterial In Vitro Techniques Luminescent Proteins/genetics Operon Promoter Regions, Genetic RNA, Messenger/genetics Receptors, Cyclic AMP/metabolism Recombinant Proteins Regulatory Sequences, Nucleic Acid Repressor Proteins Trans-Activators Vibrio/genetics
Chemicals
Bacterial Proteins DNA-Binding Proteins Luminescent Proteins RNA, Messenger Receptors, Cyclic AMP Recombinant Proteins Repressor Proteins Trans-Activators LuxR autoinducer binding proteins DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Stevens A M
Department of Microbiology, University of Iowa, Iowa City 52242.
Dolan K M
Greenberg E P
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33 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.26.12619
Published
1994-12-20
Pages
12619-23
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC45490
Subset
IM
Grants
NIAID NIH HHS · T32-AI07343 · United States
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1073/pnas.91.26.12619 PMC Full Text View journal details
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