Data represent the mean values from triplicate experiments. Discussion The results presented herein demonstrate that YmdB is a major regulator selleck inhibitor of RNase III activity in E. coli, modulating more than 30% of the genes targeted by RNase III. In addition, the results of a microarray analysis following YmdB overexpression (which identified changes in biofilm-related genes and a decrease in biofilm formation) indicate a novel role for YmdB as a modulator of biofilm formation. Previous results indicated that overexpression of RpoS was associated with decreased biofilm formation [25]. Our microarray, qPCR, and Western blotting data showed that overexpression of YmdB increased the levels of RpoS (Additional file

1: Tables S3, Figures 2, 3 and 4). Moreover, YmdB modulated RpoS levels and activity of biofilm formation (Figures 3, 4). Thus, we propose a model to illustrate the multiple roles played by YmdB during gene expression and biofilm formation (Figure 5). Figure 5 A schematic model of biofilm formation and gene expression involving YmdB, RpoS, and RNase III . Two different pathways for biofilm formation are proposed: an RNase III-dependent pathway in which other uncharacterized factor(s) inhibit RNase III activity, thereby EX 527 purchase upregulating biofilm formation, and an RNase III-independent pathway in which both YmdB and RpoS interdependently

regulate the inhibition of biofilm formation. In terms of gene expression, the level of RpoS is post-transcriptionally regulated by YmdB either

directly or indirectly via the inhibition of RNase III activity [18, 20], while the level of YmdB is regulated transcriptionally by the RpoS protein [18]. The 5′ UTR of rpoS mRNA is a known target of RNase III and its levels increase when RNase III activity is ablated [21]. Because biofilm formation is influenced by RpoS levels, it may be proposed that the rpoS mRNA is responsive to YmdB-directed RNase III inhibition. However, this is not the case because the decrease in biofilm formation following YmdB expression was not reversed in the absence of RNase III (Figure 2), suggesting that regulation of RNase III activity by YmdB is not essential for the inhibition Paclitaxel purchase of biofilm formation. Thus, the major mechanism underlying biofilm regulation by YmdB appears to be RNase III-independent (Figure 5). A screen of potential regulatory gene(s) with a YmdB-mediated phenotype demonstrated that RpoS is necessary for inhibiting biofilm formation (Figure 3); RpoS activates the transcription of ymdB[18]; thus, it is highly plausible that the RpoS gene is an upstream regulator of YmdB transcription and the resultant phenotypes. Conversely, the possibility that YmdB is a transcription factor that activates rpoS transcription was initially suggested by observations that RpoS levels were increased by YmdB overexpression, and that YmdB and RpoS are both required for the decrease in biofilm formation.