In fact, it was included as such in the Signaling Census database [28, 29]. Although sensory domains of histidine kinases are extremely

diverse, members of the same family domain typically recognize the same (or very close) substrates [39]. Therefore, we anticipated that the analysis of the two sensory domains in our histidine kinase could help us to predict its putative function. The first one showed homology to transmembrane sensory domains like PutP (Na+/proline symporter-like, in COG Evofosfamide research buy database) and SSF (sodium/solute symporter family, in Pfam database). It was preceded by a signal peptide and predicted to form twelve transmembrane helices. The second one, predicted to be cytoplasmatic, showed two PAS subdomains followed by a C-terminal PAC subdomain. In summary, the putative cognate histidine kinase of EupR was predicted to be a hybrid histidine kinase with both transmembrane and cytoplasmic sensor domains, suggesting that it could sense both external and internal conditions, and integrate them. Moreover, our in silico analysis supports the hypothesis that it may be the sensor partner of EupR. Discussion In this work, we have characterized the Tn1732-induced OSI-906 purchase salt-sensitive mutant CHR95 of C. salexigens, which showed a multiple affected phenotype: (i) inability to grow with glucose at high salinity, but not affection in the synthesis of compatible solutes, (ii) a slow growth with glucose at low and

optimal salinity, (iii) a reduced uptake and metabolism of glucose, (iv) a deregulated ectoine uptake at any salinity, and specially at low salinity, but unaffected ectoine metabolism, and (v) sensitivity to manganese.

This pleiotropic phenotype was due to deletion of three genes by the insertion of Tn1732, acs, encoding a putative acetyl-CoA synthase, mntR, encoding a manganese-dependent transcriptional Ibrutinib in vivo regulator of the DtxR/MntR family, and eupR, encoding a two-component response regulator of the NarL/FixJ family of transcriptional regulators. Transposon Tn1732 is a derivative of Tn1721, which in turn is a member of the Tn21 subgroup of the Tn3 family [40]. It has been widely used for generalized insertion mutagenesis in strains of Halomonas and selleck inhibitor Chromohalobacter, yielding single mutants [41]. However, as any Tn1721-derivative, it may cause deletions and inversions [42]. Thus, deletion of the region comprising acs-eupR-mntR upon Tn1732 insertion in CHR95 is not surprising. In fact, in the same mutagenesis experiment in which CHR95 was isolated, we also isolated the salt-sensitive mutant CHR62, showing a deletion of the ectABC genes [21, 22]. Whereas the sensitivity of strain CHR95 to manganese was correlated with the absence of mntR, its inability to grow with glucose at high salt, and the reduced transport and metabolism of glucose at low and optimal salinity (leading to a slow growth with this carbon source) may be related to deletion of the acs and/or eupR genes.