Abstract
A las-like quorum-sensing system in Pseudomonas sp. M18 was identified, which consisted of lasI and lasR genes encoding LuxI-LuxR type regulator. Several functions of the las system from strain M18 were investigated in this study. The chromosomal inactivation of either lasI or lasR by recombination increased the production of both pyoluteorin (Plt) and phenazine-1-carboxylic acid (PCA) by 4–5 fold and 2–3 fold over that of the wild type strain of M18, respectively. Production of both antibiotics was restored to wild-type levels after in trans complementation with the wild-type lasI or lasR gene. Expression of the translational fusions pltA′-′lacZ and phzA′-′lacZ further confirmed the negative effect of lasI or lasR on both biosynthetic operons, and it was also demonstrated that the las system was related to the ability of swarming motility and the inhibition of cell growth.
Similar content being viewed by others
References
Fuqua W C, Winans S C, Greenberg E P. Census and consensus in bacterial ecosystems: The Lux-LuxI family of quorum sensing transcriptional regulators. Annu Rev Microbiol, 1996, 50: 727–751
Eberl L. N-Acyl homoserine lactone-mediated gene regulation in Gram-negative bacteria. Syst Appl Microbiol, 1999, 22: 493–506
Brint J M, Ohman D E. Synthesis of multiple exoproducts in Pseudomonas aeruginosa is under the control of RhlR-RhlI, another set of regulators in strain PAO1 with homology to the autoinducer responsive LuxR-LuxI family. J Bacteriol, 1995, 177: 7155–7163
Pesci E C, Pearson J P, Seed P C, et al. Regulation of las-like and rhl quorum sensing in Pseudomonas aeruginosa. J Bacteriol, 1997, 179: 3127–3132
Schuster M, Lostroh C P, Ogi T, et al. Identification, timing, and signal specificity of Pseudomonas aeruginosa quorum-controlled genes: a transcriptome analysis. J Bacteriol, 2003, 185: 2066–2079
Chin-A-Woeng, T F C, van den Broek D, et al. Phenazine-1-carboxamide production in the biocontrol strain Pseudomonas chlororaphis PCL1391 is regulated by multiple factors secreted into the growth medium. Mol Plant Microbe Interact, 2001, 14: 969–979
Khan S R, Mavrodi D V, Jog G J, et al. Activation of the phz operon of Pseudomonas fluorescens 2-79 requires the LuxR homolog PhzR, N-(3-OHhexanoyl)-L-homoserine lactone produced by the LuxI homolog PhzI, and a cis-acting phz box. J Bacteriol, 2005, 187: 6517–6527
Wood D W, Gong F, Daykin M M, et al. N-acyl-homoserine lactone-mediated regulation of phenazine gene expression by Pseudomonas aureofaciens 30-84 in the wheat rhizosphere. J Bacteriol, 1997, 179: 7663–7670
Hu H, Xu Y, Cheng F, Zhang X. Isolation and characterization of a new fluorescent Pseudomonas strain produced both phenazine 1-carboxylic acid and pyoluteorin. J Microbiol Biotechnol, 2005, 15: 86–90
Huang X, Zhu D, Ge Y, et al. Identification and characterization of pltZ, a gene involved in the repression of pyoluteorin biosynthesis in Pseudomonas sp. M18. FEMS Microbiol Lett, 2004, 232: 197–202
Ge Y H, Huang X Q, Wang S L, et al. Phenazine-1-carboxylic acid is negatively regulated and pyoluteorin positively regulated by gacA in Pseudomonas sp. M18. FEMS Microbiol Lett, 2004, 237: 41–47
Yan A, Huang X Q, Liu H M, et al. An rhl-like quorum-sensing system negatively regulates pyoluteorin production in Pseudomonas sp. M18. Microbiology, 2007, 153: 16–28
Ge Y H, Yang S D, Yan Y, Fang Y Y, et al. RpoS as an intermediate in RsmA-dependent regulation of secondary antifungal metabolites biosynthesis in Pseudomonas sp. M18. FEMS Microbiol Lett, 2007, 268: 81–87
Hoang T T, Karkhoff-Schweizer R R, Kutchma A J, et al. A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally located DNA sequences: Application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene, 1998, 212: 77–86
Keen N T, Tamaki S, Kobayashi D, et al. Improved broad-host-range plasmids for DNA cloning in Gramnegative bacteria. Gene, 1988, 70: 191–197
Heeb S, Blumer C, Haas D. Regulatory RNA as mediator in GacA/RsmA-dependent global control of exoproduct formation in Pseudomonas fluorescens CHA0. J Bacteriol, 2002, 184: 1046–1056
Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 1989
King E O, Ward M K, Raney D E. Two simple media for the demonstration of pyocyanin and fluorescein. J Lab Clin Med, 1954, 44: 301–307
Levitch M E and Stadtman E R. A study of the biosynthesis of phenazine-1-carboxylic acid. Arch Biochem Biophys, 1964, 106: 194–199
Chen W P, Kuo T T. A simple and rapid method for the preparation of Gram-negative genomic DNA. Nucleic Acids Res, 1993, 21: 2260–2261
Sambrook J, Russell D W. Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 2001
Harunur R M, Arthur K. Inorganic polyphosphate is needed for swimming, swarming, and twitching motilities of Pseudomonas aeruginosa. Proc Natl Acad Sci USA, 2000, 97: 4885–4890
Zhang X H, Wang S L, Gen H F, et al. Differential regulation of rsmA gene on biosynthesis of pyoluteorin and phenazine-1-carboxylic acid in Pseudomonas sp. M18. World J Microbiol Biotechnol, 2005, 21: 883–889
Wang Y, Yan A, Huang X Q, et al. Construction of qscR mutant and its regulation on antibiotics production in Pseudomonas sp. M18. Acta Microbiol Sin (in Chinese), 2007, 47(2): 254–259
Yan A, Wang X L, Zhang X H, et al. LysR family factor PltR positively regulates Pyoluteorin production in a pathway-specific manner in Pseudomonas sp. M18. Sci China Ser C-Life Sci, 2007, 50(2):1–7
Brodhagen M, Henkels M D, Loper J E. Positive autoregulation and signaling properties of pyoluteorin, an antibiotic produced by the biological control organism Pseudomonas fluorescens Pf-5. Appl Environ Microbio, 2004, l 70: 1758–1766
Karin H, Vale’rie D, Marisa H, et al. Quorum-sensing-negative (lasR) mutants of Pseudomonas aeruginosa avoid cell lysis and death. J Bacteriol, 2005, 14: 4875–4883
Ruth D, Sven R, Hans H, et al. Quorum signal molecules as biosurfactants affecting swarming in Rhizobium etli. Proc Natl Acad Sci USA, 2006, 103: 14965–14970
Atkinson S, Chang C Y, Sockett E, et al. Quorum sensing in Yersinia enterocolitica controls swimming and swarming motility. J Bacteriol, 2006, 188: 1451–1461
Thilo K, Lasta K C, Francisco B, et al. Swarming of Pseudomonas aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. J Bacteriol, 2000, 182: 5990–5996
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by the Science Program of the China National 10th “Five-year Plan” (Grant No. 2004BAAA308A21-6), the National Natural Science Foundation of China (Grant No. 3037004), the National High Technology Research and Development Program (863 Program) of China (Grant No. 2007AA02Z215), New Century Excellent Talent Program (NCET) in China and Shanghai Leading Academic Discipline (Project Number: B203)
Rights and permissions
About this article
Cite this article
Chen, Y., Wang, X., Huang, X. et al. Las-like quorum-sensing system negatively regulates both pyoluteorin and phenazine-1-carboxylic acid production in Pseudomonas sp. M18. Sci. China Ser. C-Life Sci. 51, 174–181 (2008). https://doi.org/10.1007/s11427-008-0026-8
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11427-008-0026-8