Structures upstream tyrS represent the stems I, II, III and terminator of the leader region. The terminator/antiterminator
mechanism that regulates the tyrS gene is also indicated: readthrough of the leader region is induced by limitation of tyrosine. Uncharged tyrtRNA stabilize formation of antiterminator structure in the mRNA, which prevents terminator formation (SD: Shine-Dalgarno; ORF: open reading frame of tyrS) Computational three-dimensional modelling of E. durans TyrS protein revealed nucleic acids-binding domains that might suggest a role as transcriptional regulator. However, the same domains have been identified in the highly similar TyrS structure of Thermus thermophilus (Protein Data Bank: 1H3E), and predicted to interact with tRNA (Figure 6). This data is consistent with the electrophoretic mobility shift (EMSA) assays carried to test TyrS binding to AZD1208 chemical structure the promoters of the TDC operon. Under the wide range of conditions studied (different pH, salt concentration, presence or absence of tyrosine…) no specific binding of TyrS was observed (data not shown). These data, together with the finding of tyramine clusters without a tyrS gene in Tetragenococcus halophilus
(GenBank AB059363) and histamine biosynthesis clusters without a hisS gene , would suggest a non critical biological function of these genes in the modulation of the contiguous decarboxylation operon. In any case, it can not be discarded that tyrS could exert a post-transcriptional regulation of tyramine biosynthesis. In fact, both enzymes -TyrS learn more and TdcA- share tyrosine as substrate. Figure 6 TyrS structural model achieved using Swiss-Pdb Viewer v. 4.04 software and structure superposition onto the highly similar Thermus terhmophilus tyrosyl-tRNA synthetase. (Protein Data Bank: 1H3E). 1H3E is shown in green, and TyrS model is shown in magenta and yellow. 17-DMAG (Alvespimycin) HCl Analysis of the two aligned structures indicates that all of the DNA/RNA binding
sites are in regions that interact with tRNA in the 1H3E structure (shown in blue). Consequently, two are the possibilities that can be considered: i) there are two tyrS genes in E. durans -as described for E. faecalis- and the one ligated to TDC would be a stress-related gene to ensure sufficient charged TyrtRNA for protein biosynthesis in those conditions that tyrosine is being decarboxylated, or ii) this is the unique tyrS gene and the low expression levels observed under neutral pH conditions are enough to assure protein synthesis for general metabolism and the increased expression at acidic pH would guarantee protein biosynthesis when tyrosine is being decarboxylated. The presence of a second tyrS gene was investigated by Southern hybridizations of E.