Together with Cj1199 (6 2-fold), Cj1200 (14 8-fold), and Cj1422c

Together with Cj1199 (6.2-fold), Cj1200 (14.8-fold), and Cj1422c (9.1-fold) this was one of the most substantial changes observed under these conditions. Interestingly, in MHB the largest changes in transcript abundance were observed for several putative

stress response genes, which were all down-regulated in theluxSmutant. These include the putativehrcA-grpE-dnaKoperon (Cj0757-Cj0758-Cj0759; 34.1, 28.7, and 21-fold changes, respectively), and aclpBchaperone homologue (Cj0509c; 28.1-fold). Smaller changes were also observed for the putative heat shock regulatorhspR(Cj1230; 3.5-fold),crpA(Cj1229, encoding adnaJlike protein; 4-fold) and thegroES-groELoperon (Cj1220-Cj1221; 2.4 and 5.6-fold, respectively). Of these, onlyclpBtranscript levels were also changed in MEM-α (2.4-fold). Transcript changes in MHB were also observed for the putative metabolic genes Cj1364 (fumC; 10.4-fold) and Cj0481 (a putative class I aldolase; 12.1-fold), as well as the conserved hypothetical LXH254 Cj1631c (16.7-fold). For theC. jejuni luxSmutant, reduced motility Alisertib manufacturer in MHB agar plates

has been reported [35], a phenotype that was also confirmed in this study (data not shown). In agreement with these data, a set of 14 genes involved in flagella assembly and modification was found to be down-regulated in the MHB-grownluxSmutant. This includedflaA(4.2 fold lower) reported previously to be reduced in aluxSmutant of strain 81116 [44]. Interestingly, theluxSmutant was also less motile in MEM-α based motility agar, although none of the flagellar genes differentially expressed in MHB were significantly altered. However in MEM-α the transcript levels of two different putative flagellar genes Cj0336c Orotic acid (motB) and Cj1312 were significantly reduced. Two genes whose functions are associated with the AMC were found to be differentially regulated. In MHB, a 2.6-fold reduction of thepfs(Cj0117) transcript level was observed (Pfs is responsible for providing the LuxS substrate SRH), whereas in MEM-α the putativemetF(Cj1202) gene was found to be down-regulated (2.4-fold). Transcriptional changes imposed

by mutation ofluxSare not caused by a lack of AI-2-dependent signalling To test the hypothesis that a lack of extracellular AI-2 was responsible for the observed changes in the BKM120 in vitro LuxS01 transcriptome,in vitro-synthesized AI-2 was added toC. jejunicultures. The amount of AI-2 added was adjusted so that the resulting AI-2 activity at the time point of cell harvest was comparable to that produced naturally by the wild type in MHB [see Figure1]. In the case of the LuxS01 mutant,in vitrosynthesized AI-2 was added to both MEM-α and MHB grown cultures after 2.5 h. As AI-2 was not produced by the parent strain in MEM-α, it was also added after 2.5 h to test whether gene expression would be affected by quorum signalling. Levels of AI-2 in the culture supernatant were measured immediately after addition (time 0) and then again after incubation for 3.5 h and 5.5 h.

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