Structure/function Analysis of the Pantoea Stewartii Quorum-sensing Regulator EsaR As an Activator of Transcription

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2009 Oct 13

PMID 19820098

Citations 21

Authors

Daniel J Schu

Aurelien L Carlier

Katherine P Jamison

Susanne von Bodman

Ann M Stevens

Affiliations

Soon will be listed here.

Abstract

In Pantoea stewartii subsp. stewartii, two regulatory proteins are key to the process of cell-cell communication known as quorum sensing: the LuxI and LuxR homologues EsaI and EsaR. Most LuxR homologues function as activators of transcription in the presence of their cognate acylated homoserine lactone (AHL) signal. However, EsaR was initially found to function as a repressor in the absence of AHL. Previous studies demonstrated that, in the absence of AHL, EsaR retains the ability to function as a weak activator of the lux operon in recombinant Escherichia coli. Here it is shown that both the N-terminal and the C-terminal domains of EsaR are necessary for positive regulation. A site-directed mutagenesis study, guided by homology modeling to LuxR and TraR, has revealed three critical residues in EsaR that are involved in activation of RNA polymerase. In addition, a native EsaR-activated promoter has been identified, which controls expression of a putative regulatory sRNA in P. stewartii.

Citing Articles

CerM and Its Antagonist CerN Are New Components of the Quorum Sensing System in Cereibacter sphaeroides, Signaling to the CckA/ChpT/CtrA System.

Hernandez-Valle J, Vega-Baray B, Poggio S, Camarena L Microbiologyopen. 2024; 13(6):e012.

PMID: 39696824 PMC: 11655674. DOI: 10.1002/mbo3.70012.

Engineering quorum sensing-based genetic circuits enhances growth and productivity robustness of industrial E. coli at low pH.

Yan X, Bu A, Yuan Y, Zhang X, Lin Z, Yang X Microb Cell Fact. 2024; 23(1):256.

PMID: 39342182 PMC: 11438209. DOI: 10.1186/s12934-024-02524-9.

Delaying production with prokaryotic inducible expression systems.

De Baets J, De Paepe B, De Mey M Microb Cell Fact. 2024; 23(1):249.

PMID: 39272067 PMC: 11401332. DOI: 10.1186/s12934-024-02523-w.

PqsE Expands and Differentially Modulates the RhlR Quorum Sensing Regulon in Pseudomonas aeruginosa.

Letizia M, Mellini M, Fortuna A, Visca P, Imperi F, Leoni L Microbiol Spectr. 2022; 10(3):e0096122.

PMID: 35604161 PMC: 9241726. DOI: 10.1128/spectrum.00961-22.

Zinc Finger Artificial Transcription Factor-Mediated Chloroplast Genome Interrogation in Arabidopsis thaliana.

van Tol N, Flores Andaluz G, Leeggangers H, Roushan M, Hooykaas P, van der Zaal B Plant Cell Physiol. 2018; 60(2):393-406.

PMID: 30398644 PMC: 6375250. DOI: 10.1093/pcp/pcy216.

References

Higuchi R, Krummel B, Saiki R . A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988; 16(15):7351-67. PMC: 338413. DOI: 10.1093/nar/16.15.7351. View

Qin Y, Luo Z, Farrand S . Domains formed within the N-terminal region of the quorum-sensing activator TraR are required for transcriptional activation and direct interaction with RpoA from agrobacterium. J Biol Chem. 2004; 279(39):40844-51. DOI: 10.1074/jbc.M405299200. View

Miller M, Bassler B . Quorum sensing in bacteria. Annu Rev Microbiol. 2001; 55:165-99. DOI: 10.1146/annurev.micro.55.1.165. View

White C, Winans S . Identification of amino acid residues of the Agrobacterium tumefaciens quorum-sensing regulator TraR that are critical for positive control of transcription. Mol Microbiol. 2005; 55(5):1473-86. DOI: 10.1111/j.1365-2958.2004.04482.x. View

Carlier A, von Bodman S . The rcsA promoter of Pantoea stewartii subsp. stewartii features a low-level constitutive promoter and an EsaR quorum-sensing-regulated promoter. J Bacteriol. 2006; 188(12):4581-4. PMC: 1482964. DOI: 10.1128/JB.00211-06. View

Dykxhoorn D, St Pierre R, Linn T . A set of compatible tac promoter expression vectors. Gene. 1996; 177(1-2):133-6. DOI: 10.1016/0378-1119(96)00289-2. View

Grant S, Jessee J, Bloom F, Hanahan D . Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants. Proc Natl Acad Sci U S A. 1990; 87(12):4645-9. PMC: 54173. DOI: 10.1073/pnas.87.12.4645. View

Atkinson S, Chang C, Sockett R, Camara M, Williams P . Quorum sensing in Yersinia enterocolitica controls swimming and swarming motility. J Bacteriol. 2006; 188(4):1451-61. PMC: 1367215. DOI: 10.1128/JB.188.4.1451-1461.2006. View

Beck von Bodman S, Farrand S . Capsular polysaccharide biosynthesis and pathogenicity in Erwinia stewartii require induction by an N-acylhomoserine lactone autoinducer. J Bacteriol. 1995; 177(17):5000-8. PMC: 177277. DOI: 10.1128/jb.177.17.5000-5008.1995. View

10.

Hanahan D . Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983; 166(4):557-80. DOI: 10.1016/s0022-2836(83)80284-8. View

11.

De Lay N, Gottesman S . The Crp-activated small noncoding regulatory RNA CyaR (RyeE) links nutritional status to group behavior. J Bacteriol. 2008; 191(2):461-76. PMC: 2620814. DOI: 10.1128/JB.01157-08. View

12.

Vanderpool C, Gottesman S . The novel transcription factor SgrR coordinates the response to glucose-phosphate stress. J Bacteriol. 2007; 189(6):2238-48. PMC: 1899371. DOI: 10.1128/JB.01689-06. View

13.

Johnson D, Ishihama A, Stevens A . Involvement of region 4 of the sigma70 subunit of RNA polymerase in transcriptional activation of the lux operon during quorum sensing. FEMS Microbiol Lett. 2003; 228(2):193-201. DOI: 10.1016/S0378-1097(03)00750-X. View

14.

Qin Y, Luo Z, Smyth A, Gao P, Beck von Bodman S, Farrand S . Quorum-sensing signal binding results in dimerization of TraR and its release from membranes into the cytoplasm. EMBO J. 2000; 19(19):5212-21. PMC: 302097. DOI: 10.1093/emboj/19.19.5212. View

15.

Morohoshi T, Nakamura Y, Yamazaki G, Ishida A, Kato N, Ikeda T . The plant pathogen Pantoea ananatis produces N-acylhomoserine lactone and causes center rot disease of onion by quorum sensing. J Bacteriol. 2007; 189(22):8333-8. PMC: 2168703. DOI: 10.1128/JB.01054-07. View

16.

Whitehead N, Barnard A, Slater H, Simpson N, Salmond G . Quorum-sensing in Gram-negative bacteria. FEMS Microbiol Rev. 2001; 25(4):365-404. DOI: 10.1111/j.1574-6976.2001.tb00583.x. View

17.

Luo Z, Farrand S . Signal-dependent DNA binding and functional domains of the quorum-sensing activator TraR as identified by repressor activity. Proc Natl Acad Sci U S A. 1999; 96(16):9009-14. PMC: 17723. DOI: 10.1073/pnas.96.16.9009. View

18.

Fuqua C, Parsek M, Greenberg E . Regulation of gene expression by cell-to-cell communication: acyl-homoserine lactone quorum sensing. Annu Rev Genet. 2001; 35:439-68. DOI: 10.1146/annurev.genet.35.102401.090913. View

19.

Minogue T, Bernhard F, von Bodman S . The autoregulatory role of EsaR, a quorum-sensing regulator in Pantoea stewartii ssp. stewartii: evidence for a repressor function. Mol Microbiol. 2002; 44(6):1625-35. DOI: 10.1046/j.1365-2958.2002.02987.x. View

20.

Withers H, Swift S, Williams P . Quorum sensing as an integral component of gene regulatory networks in Gram-negative bacteria. Curr Opin Microbiol. 2001; 4(2):186-93. DOI: 10.1016/s1369-5274(00)00187-9. View