4). The last two results suggested a σE-dependent regulation of the sbmA promoter. In contrast to the above results, eliminating σE

would reduce the expression of sbmA. Although rpoE is essential, it could be deleted from the strain SC122 (Rouviere et al., 1995) in the presence of an uncharacterized suppressor mutation (Alba et al., 2001), obtaining the CAG22222 strain. This allows a comparison of the specific activity of the ΔsbmA∷lacZY fusion (transduced in the two above-mentioned strains) in the presence and absence of σE. The stationary-phase activity of ΔsbmA∷lacZY fusion seen in the wild-type rpoE+ context (NC122 strain) was almost completely abolished in an rpoE background (NC322 strain) (Fig. 5). On the other hand, the induction of the ΔsbmA∷lacZY fusion activity by ethanol addition was also observed in the NC122 fusion strain and was reverted in the absence of rpoE (NC322 strain) click here (data not shown). The last result confirms the σE-mediated induction of sbmA by this extracytoplasmatic stress. In order to evaluate the influence Crizotinib of σE on the tolC mutation-dependent upregulation

of sbmA, the tolC rpoE double mutant of the ΔsbmA∷lacZY fusion was constructed. To this end, a P1 transduction was performed with a tolC∷Tn10 mutant, as a donor, and NC122 and NC322 strains, as recipients, obtaining the NC222 and NC422 strains, respectively. Figure 5 shows that the increase in the β-galactosidase activity of ΔsbmA∷lacZY fusion produced in a tolC context (NC222 strain) disappears when rpoE is absent (NC422 strain). Altogether, these PTK6 results strongly support the idea that the transcriptional induction of sbmA by tolC mutation is completely σE dependent. It is well known that tolC mutants are pleiotropic and extremely sensitive to detergents and dyes, mainly due to the inability to pump out noxious compounds. In this mutant, a membrane permeability defect

was also demonstrated that involved a modification in the OmpF/OmpC ratio, pushing this in favor of OmpC (and MicF) (Misra & Reeves, 1987). Recently, we demonstrated that the tolC mutation severely reduces high-level resistance to tetracycline (de Cristobal et al., 2008). These results have indicated that TolC is critical for E. coli survival in an environment with noxious compounds. We also found that the inactivation of sbmA alone partially inhibited high-level tetracycline resistance. Moreover, the sbmA tolC double mutation had an additive effect, resulting in almost complete suppression of the phenotypic expression of Tn10 tetracycline resistance. In this paper, we showed that there is an sbmA-positive regulation in response to the absence of tolC, mediated by σE activation. This upregulation could be caused by the alteration in outer membrane permeability.