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Functional domains of the RhlR transcriptional regulator of Pseudomonas aeruginosa.
J Bacteriol. 2003 Dec; 185(24):7129-39.JB

Abstract

The RhlR transcriptional regulator of Pseudomonas aeruginosa, along with its cognate autoinducer, N-butyryl homoserine lactone (C(4)-HSL), regulates gene expression in response to cell density. With an Escherichia coli LexA-based protein interaction system, we demonstrated that RhlR multimerized and that the degree of multimerization was dependent on the C(4)-HSL concentration. Studies with an E. coli lasB::lacZ lysogen demonstrated that RhlR multimerization was necessary for it to function as a transcriptional activator. Deletion analysis of RhlR indicated that the N-terminal domain of the protein is necessary for C(4)-HSL binding. Single amino acid substitutions in the C-terminal domain of RhlR generated mutant RhlR proteins that had the ability to bind C(4)-HSL and multimerize but were unable to activate lasB expression, demonstrating that the C-terminal domain is important for target gene activation. Single amino acid substitutions in both the N-terminal and C-terminal domains of RhlR demonstrated that both domains possess residues involved in multimerization. RhlR with a C-terminal deletion and an RhlR site-specific mutant form that possessed multimerization but not transcriptional activation capabilities were able to inhibit the ability of wild-type RhlR to activate rhlA expression in P. aeruginosa. We conclude that C(4)-HSL binding is necessary for RhlR multimerization and that RhlR functions as a multimer in P. aeruginosa.

Authors+Show Affiliations

University of Rochester, Rochester, New York 14642, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

14645272

Citation

Lamb, Janet R., et al. "Functional Domains of the RhlR Transcriptional Regulator of Pseudomonas Aeruginosa." Journal of Bacteriology, vol. 185, no. 24, 2003, pp. 7129-39.
Lamb JR, Patel H, Montminy T, et al. Functional domains of the RhlR transcriptional regulator of Pseudomonas aeruginosa. J Bacteriol. 2003;185(24):7129-39.
Lamb, J. R., Patel, H., Montminy, T., Wagner, V. E., & Iglewski, B. H. (2003). Functional domains of the RhlR transcriptional regulator of Pseudomonas aeruginosa. Journal of Bacteriology, 185(24), 7129-39.
Lamb JR, et al. Functional Domains of the RhlR Transcriptional Regulator of Pseudomonas Aeruginosa. J Bacteriol. 2003;185(24):7129-39. PubMed PMID: 14645272.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional domains of the RhlR transcriptional regulator of Pseudomonas aeruginosa. AU - Lamb,Janet R, AU - Patel,Hetal, AU - Montminy,Timothy, AU - Wagner,Victoria E, AU - Iglewski,Barbara H, PY - 2003/12/4/pubmed PY - 2004/1/8/medline PY - 2003/12/4/entrez SP - 7129 EP - 39 JF - Journal of bacteriology JO - J. Bacteriol. VL - 185 IS - 24 N2 - The RhlR transcriptional regulator of Pseudomonas aeruginosa, along with its cognate autoinducer, N-butyryl homoserine lactone (C(4)-HSL), regulates gene expression in response to cell density. With an Escherichia coli LexA-based protein interaction system, we demonstrated that RhlR multimerized and that the degree of multimerization was dependent on the C(4)-HSL concentration. Studies with an E. coli lasB::lacZ lysogen demonstrated that RhlR multimerization was necessary for it to function as a transcriptional activator. Deletion analysis of RhlR indicated that the N-terminal domain of the protein is necessary for C(4)-HSL binding. Single amino acid substitutions in the C-terminal domain of RhlR generated mutant RhlR proteins that had the ability to bind C(4)-HSL and multimerize but were unable to activate lasB expression, demonstrating that the C-terminal domain is important for target gene activation. Single amino acid substitutions in both the N-terminal and C-terminal domains of RhlR demonstrated that both domains possess residues involved in multimerization. RhlR with a C-terminal deletion and an RhlR site-specific mutant form that possessed multimerization but not transcriptional activation capabilities were able to inhibit the ability of wild-type RhlR to activate rhlA expression in P. aeruginosa. We conclude that C(4)-HSL binding is necessary for RhlR multimerization and that RhlR functions as a multimer in P. aeruginosa. SN - 0021-9193 UR - https://www.unboundmedicine.com/medline/citation/14645272/Functional_domains_of_the_RhlR_transcriptional_regulator_of_Pseudomonas_aeruginosa_ L2 - http://jb.asm.org/cgi/pmidlookup?view=long&pmid=14645272 DB - PRIME DP - Unbound Medicine ER -