{"refrec":{"BRefID":114716,"RR":"<b>Defoirdt, T.; Miyamoto, C.M.; Wood, T.K.; Meighen, E.A.; Sorgeloos, P.; Verstraete, W.; Bossier, P.</b> (2007). The natural furanone (5<i>Z</i>)-4-bromo-5-(bromomethylene)-3-butyl-2(5<i>H</i>)-furanone disrupts quorum sensing-regulated gene expression in <i>Vibrio harveyi</i> by decreasing the DNA-binding activity of the transcriptional regulator protein luxR. <i>Environ. Microbiol. 9(10)</i>: 2486-2495. <a href=\"http://dx.doi.org/10.1111/j.1462-2920.2007.01367.x\" target=\"_blank\">dx.doi.org/10.1111/j.1462-2920.2007.01367.x</a>","BEntID":109111,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":1,"RefStringPartII":". <i>Environ. Microbiol. 9(10)</i>: 2486-2495. <a href=\"https://dx.doi.org/10.1111/j.1462-2920.2007.01367.x\" target=\"_blank\">https://dx.doi.org/10.1111/j.1462-2920.2007.01367.x</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Defoirdt, T.; Miyamoto, C.M.; Wood, T.K.; Meighen, E.A.; Sorgeloos, P.; Verstraete, W.; Bossier, P.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Defoirdt, T. <i>et al.</i>","Englishabstract":"This study aimed at getting a deeper insight in the molecular mechanism by which the natural furanone (5<i>Z</i>)-4-bromo-5-(bromomethylene)-3-butyl-2(5<i>H</i>)- furanone disrupts quorum sensing in Vibrio harveyi. Bioluminescence experiments with signal molecule receptor double mutants revealed that the furanone blocks all three channels of the <i>V. harveyi</i> quorum sensing system. In further experiments using mutants with mutations in the quorum sensing signal transduction pathway, the compound was found to block quorum sensing-regulated bioluminescence by interacting with a component located downstream of the Hfq protein. Furthermore, reverse transcriptase real-time polymerase chain reaction with specific primers showed that there was no effect of the furanone on luxR<sub>Vh</sub> mRNA levels in wild-type <i>V. harveyi</i> cells. In contrast, mobility shift assays showed that in the presence of the furanone, significantly lower levels of the LuxR<sub>Vh</sub> response regulator protein were able to bind to its target promoter sequences in wild-type <i>V. harveyi</i>. Finally, tests with purified LuxR<sub>Vh</sub> protein also showed less shifts with furanone-treated LuxR<sub>Vh</sub>, whereas the LuxR<sub>Vh</sub> concentration was found not to be altered by the furanone (as determined by SDS-PAGE). Therefore, our data indicate that the furanone blocks quorum sensing in <i>V. harveyi</i> by rendering the quorum sensing master regulator protein LuxR<sub>Vh</sub> unable to bind to the promoter sequences of quorum sensing-regulated genes. 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