DNA binding kinetics of two response regulators, PlnC and PlnD, from the bacteriocin regulon of Lactobacillus plantarum C11
Tóm tắt
Bacteriocin production in the lactic acid bacterium Lactobacillus plantarum C11 is regulated through a quorum sensing based pathway involving two highly homologous response regulators (59% identity and 76% similarity), PlnC as a transcriptional activator and PlnD as a repressor. Previous in vitro studies have shown that both regulators bind, as homodimers, to the same DNA regulatory repeats to exert their regulatory functions. As the genes for these two proteins are located on the same auto-regulatory operon, hence being co-expressed upon gene activation, it is plausible that their opposite functions must somehow be differentially regulated, either in terms of timing and/or binding kinetics, so that their activities do not impair each other in an uncontrolled manner. To understand the nature behind this potential differentiation, we have studied the binding kinetics of the two regulators on five target promoters (P
plnA
, P
plnM
, P
plnJ
, P
plnE
and P
plnG
) from the bacteriocin regulon of L. plantarum C11. By using surface plasmon resonance spectroscopy we obtained parameters such as association rates, dissociation rates and dissociation constants, showing that the two regulators indeed differ greatly from each other in terms of cooperative binding and binding strength to the different promoters. For instance, cooperativity is very strong for PlnC binding to the promoter of the regulatory operon (P
plnA
), but not to the promoter of the transport operon (P
plnG
), while the opposite is seen for PlnD binding to these two promoters. The estimated affinity constants indicate that PlnC can bind to P
plnA
to activate transcription of the key regulatory operon plnABCD without much interference from PlnD, and that the repressive function of PlnD might act through a different mechanism than repression of the regulatory operon. We have characterised the DNA binding kinetics of the two regulators PlnC and PlnD from the bacteriocin locus in L. plantarum C11. Our data show that PlnC and PlnD, despite their strong homology to each other, differ greatly from each other in terms of binding affinity and cooperativity to the different promoters of the pln regulon.
Tài liệu tham khảo
Moll GN, Akker van den E, Hauge HH, Nissen-Meyer J, Nes IF, Konings WN, Driessen AJ: Complementary and overlapping selectivity of the two-peptide bacteriocins plantaricin EF and JK. J Bacteriol. 1999, 181 (16): 4848-4852.
Cascales E, Buchanan SK, Duche D, Kleanthous C, Lloubes R, Postle K, Riley M, Slatin S, Cavard D: Colicin biology. Microbiol Mol Biol Rev. 2007, 71 (1): 158-229. 10.1128/MMBR.00036-06.
Oppegård C, Rogne P, Emanuelsen L, Kristiansen PE, Fimland G, Nissen-Meyer J: The two-peptide class II bacteriocins: structure, production, and mode of action. J Mol Microbiol Biotechnol. 2007, 13 (4): 210-219. 10.1159/000104750.
Nes IF, Diep DB, Havarstein LS, Brurberg MB, Eijsink V, Holo H: Biosynthesis of bacteriocins in lactic acid bacteria. Antonie Van Leeuwenhoek. 1996, 70 (2–4): 113-128. 10.1007/BF00395929.
Nes IF, Eijsink VGH: Regulation of Group II Peptide Bacteriocin Synthesis by Quorum-Sensing Mechanisms. American Society for Microbiology. 1999, 175-192.
Rojo-Bezares B, Saenz Y, Navarro L, Zarazaga M, Ruiz-Larrea F, Torres C: Coculture-inducible bacteriocin activity of Lactobacillus plantarum strain J23 isolated from grape must. Food Microbiol. 2007, 24 (5): 482-491. 10.1016/j.fm.2006.09.003.
Maldonado A, Jimenez-Diaz R, Ruiz-Barba JL: Induction of plantaricin production in Lactobacillus plantarum NC8 after coculture with specific gram-positive bacteria is mediated by an autoinduction mechanism. J Bacteriol. 2004, 186 (5): 1556-1564. 10.1128/JB.186.5.1556-1564.2004.
Chang JY, Lee HJ, Chang HC: Identification of the agent from Lactobacillus plantarum KFRI464 that enhances bacteriocin production by Leuconostoc citreum GJ7. J Appl Microbiol. 2007, 103 (6): 2504-2515. 10.1111/j.1365-2672.2007.03543.x.
Diep DB, Havarstein LS, Nes IF: Characterization of the locus responsible for the bacteriocin production in Lactobacillus plantarum C11. J Bacteriol. 1996, 178 (15): 4472-4483.
Brurberg MB, Nes IF, Eijsink VG: Pheromone-induced production of antimicrobial peptides in Lactobacillus. Mol Microbiol. 1997, 26 (2): 347-360. 10.1046/j.1365-2958.1997.5821951.x.
Risøen PA, Havarstein LS, Diep DB, Nes IF: Identification of the DNA-binding sites for two response regulators involved in control of bacteriocin synthesis in Lactobacillus plantarum C11. Mol Gen Genet. 1998, 259 (2): 224-232.
Pei J, Grishin NV: Type II CAAX prenyl endopeptidases belong to a novel superfamily of putative membrane-bound metalloproteases. Trends Biochem Sci. 2001, 26 (5): 275-277. 10.1016/S0968-0004(01)01813-8.
Nikolskaya AN, Galperin MY: A novel type of conserved DNA-binding domain in the transcriptional regulators of the AlgR/AgrA/LytR family. Nucleic Acids Res. 2002, 30 (11): 2453-2459. 10.1093/nar/30.11.2453.
Sidote DJ, Barbieri CM, Wu T, Stock AM: Structure of the Staphylococcus aureus AgrA LytTR domain bound to DNA reveals a beta fold with an unusual mode of binding. Structure. 2008, 16 (5): 727-735. 10.1016/j.str.2008.02.011.
Diep DB, Johnsborg O, Risoen PA, Nes IF: Evidence for dual functionality of the operon plnABCD in the regulation of bacteriocin production in Lactobacillus plantarum. Mol Microbiol. 2001, 41 (3): 633-644. 10.1046/j.1365-2958.2001.02533.x.
Diep DB, Myhre R, Johnsborg O, Aakra A, Nes IF: Inducible bacteriocin production in Lactobacillus is regulated by differential expression of the pln operons and by two antagonizing response regulators, the activity of which is enhanced upon phosphorylation. Mol Microbiol. 2003, 47 (2): 483-494. 10.1046/j.1365-2958.2003.03310.x.
Håvarstein LS, Gaustad P, Nes IF, Morrison DA: Identification of the streptococcal competence-pheromone receptor. Mol Microbiol. 1996, 21 (4): 863-869. 10.1046/j.1365-2958.1996.521416.x.
Novick RP, Projan SJ, Kornblum J, Ross HF, Ji G, Kreiswirth B, Vandenesch F, Moghazeh S: The agr P2 operon: an autocatalytic sensory transduction system in Staphylococcus aureus. Mol Gen Genet. 1995, 248 (4): 446-458. 10.1007/BF02191645.
Hancock LE, Perego M: The Enterococcus faecalis fsr two-component system controls biofilm development through production of gelatinase. J Bacteriol. 2004, 186 (17): 5629-5639. 10.1128/JB.186.17.5629-5639.2004.
Maldonado A, Ruiz-Barba JL, Jimenez-Diaz R: Purification and genetic characterization of plantaricin NC8, a novel coculture-inducible two-peptide bacteriocin from Lactobacillus plantarum NC8. Appl Environ Microbiol. 2003, 69 (1): 383-389. 10.1128/AEM.69.1.383-389.2003.
Rojo-Bezares B, Saenz Y, Navarro L, Jimenez-Diaz R, Zarazaga M, Ruiz-Larrea F, Torres C: Characterization of a new organization of the plantaricin locus in the inducible bacteriocin-producing Lactobacillus plantarum J23 of grape must origin. Arch Microbiol. 2008, 189 (5): 491-499. 10.1007/s00203-007-0342-6.
Risøen PA, Johnsborg O, Diep DB, Hamoen L, Venema G, Nes IF: Regulation of bacteriocin production in Lactobacillus plantarum depends on a conserved promoter arrangement with consensus binding sequence. Mol Genet Genomics. 2001, 265 (1): 198-206. 10.1007/s004380000397.
Navarro L, Rojo-Bezares B, Saenz Y, Diez L, Zarazaga M, Ruiz-Larrea F, Torres C: Comparative study of the pln locus of the quorum-sensing regulated bacteriocin-producing L. plantarum J51 strain. Int J Food Microbiol. 2008, 128 (2): 390-394. 10.1016/j.ijfoodmicro.2008.08.004.
Villegas A, Kropinski AM: An analysis of initiation codon utilization in the Domain Bacteria – concerns about the quality of bacterial genome annotation. Microbiology. 2008, 154 (Pt 9): 2559-2661. 10.1099/mic.0.2008/021360-0.
Francke C, Kerkhoven R, Wels M, Siezen R: A generic approach to identify Transcription Factor-specific operator motifs; Inferences for LacI-family mediated regulation in Lactobacillus plantarum WCFS1. BMC Genomics. 2008, 9 (145):
Galperin MY: Telling bacteria: do not LytTR. Structure. 2008, 16 (5): 657-659. 10.1016/j.str.2008.04.003.
Schillinger U, Lucke FK: Antibacterial activity of Lactobacillus sake isolated from meat. Appl Environ Microbiol. 1989, 55 (8): 1901-1906.
Sørvig E, Mathiesen G, Naterstad K, Eijsink VG, Axelsson L: High-level, inducible gene expression in Lactobacillus sakei and Lactobacillus plantarum using versatile expression vectors. Microbiology. 2005, 151 (Pt 7): 2439-2449. 10.1099/mic.0.28084-0.
Straume D, Axelsson L, Nes IF, Diep DB: Improved expression and purification of the correctly folded response regulator PlnC from lactobacilli. J Microbiol Methods. 2006, 67: 191-203. 10.1016/j.mimet.2006.03.022.