Molecular Mechanisms of Master Regulator VqsM Mediating Quorum-sensing and Antibiotic Resistance in Pseudomonas Aeruginosa

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2014 Jul 19

PMID 25034696

Citations 47

Authors

Haihua Liang

Xin Deng

Xuefeng Li

Yan Ye

Min Wu

Affiliations

Soon will be listed here.

Abstract

The Pseudomonas aeruginosa quorum-sensing (QS) systems contribute to bacterial homeostasis and pathogenicity. Although the AraC-family transcription factor VqsM has been characterized to control the production of virulence factors and QS signaling molecules, its detailed regulatory mechanisms still remain elusive. Here, we report that VqsM directly binds to the lasI promoter region, and thus regulates its expression. To identify additional targets of VqsM in P. aeruginosa PAO1, we performed chromatin immunoprecipitation (ChIP) followed by high-throughput DNA sequencing (ChIP-seq) and detected 48 enriched loci harboring VqsM-binding peaks in the P. aeruginosa genome. The direct regulation of these genes by VqsM has been confirmed by electrophoretic mobility shift assays and quantitative real-time polymerase chain reactions. A VqsM-binding motif was identified by using the MEME suite and verified by footprint assays in vitro. In addition, VqsM directly bound to the promoter regions of the antibiotic resistance regulator NfxB and the master type III secretion system (T3SS) regulator ExsA. Notably, the vqsM mutant displayed more resistance to two types of antibiotics and promoted bacterial survival in a mouse model, compared to wild-type PAO1. Collectively, this work provides new cues to better understand the detailed regulatory networks of QS systems, T3SS, and antibiotic resistance.

Citing Articles

Understanding the pathophysiology of colonization as a guide for future treatment for chronic leg ulcers.

Matheus G, Chamoun M, Khosrotehrani K, Sivakumaran Y, Wells T Burns Trauma. 2025; 13():tkae083.

PMID: 39830194 PMC: 11741523. DOI: 10.1093/burnst/tkae083.

Regulation mechanism of the long-chain -alkane monooxygenase gene in RAG-1.

Chen S, Cao L, Lv T, Liu J, Gao G, Li M Appl Environ Microbiol. 2024; 91(1):e0205024.

PMID: 39723816 PMC: 11784139. DOI: 10.1128/aem.02050-24.

An insight on the powerful of bacterial quorum sensing inhibition.

Naga N, Shaaban M, El-Metwally M Eur J Clin Microbiol Infect Dis. 2024; 43(11):2071-2081.

PMID: 39158799 PMC: 11534983. DOI: 10.1007/s10096-024-04920-w.

The quorum sensing regulator RhlR positively controls the expression of the type III secretion system in Pseudomonas aeruginosa PAO1.

Montelongo-Martinez L, Diaz-Guerrero M, Flores-Vega V, Soto-Aceves M, Rosales-Reyes R, Quiroz-Morales S PLoS One. 2024; 19(8):e0307174.

PMID: 39146292 PMC: 11326643. DOI: 10.1371/journal.pone.0307174.

Ser/Thr protein kinase Stk1 phosphorylates the key transcriptional regulator AlgR to modulate virulence and resistance in .

Li R, Zhu X, Zhang P, Wu X, Jin Q, Pan J Virulence. 2024; 15(1):2367649.

PMID: 38898809 PMC: 11197903. DOI: 10.1080/21505594.2024.2367649.

References

Schuster M, Greenberg E . A network of networks: quorum-sensing gene regulation in Pseudomonas aeruginosa. Int J Med Microbiol. 2006; 296(2-3):73-81. DOI: 10.1016/j.ijmm.2006.01.036. View

Williams P, Camara M . Quorum sensing and environmental adaptation in Pseudomonas aeruginosa: a tale of regulatory networks and multifunctional signal molecules. Curr Opin Microbiol. 2009; 12(2):182-91. DOI: 10.1016/j.mib.2009.01.005. View

Kannan S, Audet A, Knittel J, Mullegama S, Gao G, Wu M . Src kinase Lyn is crucial for Pseudomonas aeruginosa internalization into lung cells. Eur J Immunol. 2006; 36(7):1739-52. DOI: 10.1002/eji.200635973. View

Rampioni G, Bertani I, Zennaro E, Polticelli F, Venturi V, Leoni L . The quorum-sensing negative regulator RsaL of Pseudomonas aeruginosa binds to the lasI promoter. J Bacteriol. 2005; 188(2):815-9. PMC: 1347304. DOI: 10.1128/JB.188.2.815-819.2006. View

Chugani S, Whiteley M, Lee K, DArgenio D, Manoil C, Greenberg E . QscR, a modulator of quorum-sensing signal synthesis and virulence in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2001; 98(5):2752-7. PMC: 30211. DOI: 10.1073/pnas.051624298. View

Schuster M, Urbanowski M, Greenberg E . Promoter specificity in Pseudomonas aeruginosa quorum sensing revealed by DNA binding of purified LasR. Proc Natl Acad Sci U S A. 2004; 101(45):15833-9. PMC: 528741. DOI: 10.1073/pnas.0407229101. View

Pearson J, Gray K, Passador L, Tucker K, Eberhard A, Iglewski B . Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes. Proc Natl Acad Sci U S A. 1994; 91(1):197-201. PMC: 42913. DOI: 10.1073/pnas.91.1.197. View

Stols L, Gu M, Dieckman L, Raffen R, Collart F, Donnelly M . A new vector for high-throughput, ligation-independent cloning encoding a tobacco etch virus protease cleavage site. Protein Expr Purif. 2002; 25(1):8-15. DOI: 10.1006/prep.2001.1603. View

Siehnel R, Traxler B, An D, Parsek M, Schaefer A, Singh P . A unique regulator controls the activation threshold of quorum-regulated genes in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2010; 107(17):7916-21. PMC: 2867882. DOI: 10.1073/pnas.0908511107. View

10.

Bailey T, Boden M, Buske F, Frith M, Grant C, Clementi L . MEME SUITE: tools for motif discovery and searching. Nucleic Acids Res. 2009; 37(Web Server issue):W202-8. PMC: 2703892. DOI: 10.1093/nar/gkp335. View

11.

McGrath S, Wade D, Pesci E . Dueling quorum sensing systems in Pseudomonas aeruginosa control the production of the Pseudomonas quinolone signal (PQS). FEMS Microbiol Lett. 2004; 230(1):27-34. DOI: 10.1016/S0378-1097(03)00849-8. View

12.

Lee J, Wu J, Deng Y, Wang J, Wang C, Wang J . A cell-cell communication signal integrates quorum sensing and stress response. Nat Chem Biol. 2013; 9(5):339-43. DOI: 10.1038/nchembio.1225. View

13.

OToole G, Kolter R . Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol. 1998; 30(2):295-304. DOI: 10.1046/j.1365-2958.1998.01062.x. View

14.

Wurtzel O, Yoder-Himes D, Han K, Dandekar A, Edelheit S, Greenberg E . The single-nucleotide resolution transcriptome of Pseudomonas aeruginosa grown in body temperature. PLoS Pathog. 2012; 8(9):e1002945. PMC: 3460626. DOI: 10.1371/journal.ppat.1002945. View

15.

Okazaki T, Iyobe S, Hashimoto H, Hirai K . Cloning and characterization of a DNA fragment that complements the nfxB mutation in Pseudomonas aeruginosa PAO. FEMS Microbiol Lett. 1991; 63(1):31-5. DOI: 10.1016/0378-1097(91)90522-c. View

16.

Dong Y, Zhang X, Xu J, Tan A, Zhang L . VqsM, a novel AraC-type global regulator of quorum-sensing signalling and virulence in Pseudomonas aeruginosa. Mol Microbiol. 2005; 58(2):552-64. DOI: 10.1111/j.1365-2958.2005.04851.x. View

17.

Blasco B, Chen J, Hartkoorn R, Sala C, Uplekar S, Rougemont J . Virulence regulator EspR of Mycobacterium tuberculosis is a nucleoid-associated protein. PLoS Pathog. 2012; 8(3):e1002621. PMC: 3315491. DOI: 10.1371/journal.ppat.1002621. View

18.

Bokhove M, Nadal Jimenez P, Quax W, Dijkstra B . The quorum-quenching N-acyl homoserine lactone acylase PvdQ is an Ntn-hydrolase with an unusual substrate-binding pocket. Proc Natl Acad Sci U S A. 2010; 107(2):686-91. PMC: 2818923. DOI: 10.1073/pnas.0911839107. View

19.

Gallagher L, McKnight S, Kuznetsova M, Pesci E, Manoil C . Functions required for extracellular quinolone signaling by Pseudomonas aeruginosa. J Bacteriol. 2002; 184(23):6472-80. PMC: 135424. DOI: 10.1128/JB.184.23.6472-6480.2002. View

20.

Liang H, Li L, Dong Z, Surette M, Duan K . The YebC family protein PA0964 negatively regulates the Pseudomonas aeruginosa quinolone signal system and pyocyanin production. J Bacteriol. 2008; 190(18):6217-27. PMC: 2546791. DOI: 10.1128/JB.00428-08. View