1C, Graphs 2 and 3); 3) in an arabinose-inducible promoter system, production of InvE protein decreased under low osmotic conditions even in the presence of sufficient amounts of invE mRNA (Fig. 2A); 4) in the absence of the RNA chaperone Hfq, the amount of InvE protein correlated with the level of virF transcription, even in low osmotic conditions (Fig. 3A); 5) InvE production was reduced upon over-expression of Hfq protein, even in physiological osmotic conditions (Fig. 3B); and 6) the stability of invE
mRNA decreased under low osmotic conditions in the wild-type strain, but click here was increased in the hfq mutant (Fig. 4). The synthesis of TTSS is induced in response to changes in osmolarity. While several osmolytes were able to induce TTSS synthesis, the response was weaker with the non-salt osmolyte sorbitol. Differences in TTSS synthesis in response to different osmolytes might be due to differences in permeability or influx through the bacterial membrane. Under GSK872 solubility dmso physiological conditions, the contribution of non-salt osmolytes is likely to less relevant, because carbohydrates are almost completely absorbed in the ileum before reaching the colon, where infection and propagation of Shigella takes place. In the colon, Na+ ions and water are actively absorbed,
and K+ ions are passively secreted, leading to an induction of TTSS synthesis. However, we did not observe significant differences in the expression of TTSS (Fig. 1A) and invasion (data not shown) in the presence of the two ions, which indicates that the trigger for TTSS induction is ionic strength, and not the nature of the ionic species. In prokaryotes, the GSK126 in vitro regulation Cobimetinib clinical trial of gene expression takes place mainly
at the level of transcription. In the expression of a set of genes, however, regulation takes place at any one of several post-transcriptional stages, including the regulation of mRNA stability and translation, through a variety of mechanisms. We propose a model for the post-transcriptional repression of InvE expression in which the association of invE mRNA with the RNA chaperone Hfq controls mRNA stability. Recently, it was suggested that an iron-regulated small RNA, RyhB , plays a regulatory role in invE expression . At present, we cannot rule out the possibility that an interaction between invE mRNA and an as-yet unidentified RNA is involved in the temperature- and osmotic pressure-dependent activation of InvE synthesis. To date, various mechanisms have been proposed for the regulation of translation initiation through the modulation of RNA structure, including the structure of the initiation codon .