2 6E-05     cpe1386 Unknown (85aa) 8.73 < 1E-05     cpe1387 Unknown (71aa) 6.4 5E-06     cpe1388 Unknown 5.94 5E-05     cpe0651 Unknown 4.8 < 1E-05     cpe0015 Unknown 4.7 2E-05     cpe0114 Unknown (74 aa) 4.49 1.33E-05     cpe0113 Unknown (75 aa) 3.98 6E-05     cpe0264 Unknown (98 aa) 3.94 0.001     cpe2619 Unknown (63 aa) 3.88 < 1E-05     cpe0067 Unknown 3.6 1E-05     cpe0102 Unknown (90 aa) 3.52 0.01     cpe1472 Unknown 3.36 0.00735 click here     cpe2037 Unknown (89 aa) 3.16 0.0006     cpe0363 Unknown (118 aa) 3.11 0.002     The results presented are representative of 8 differential hybridizations using RNA preparations

obtained from 4 independent cultures. aa means amino acids. Regulation of T-box controlled genes Among genes derepressed after growth in the presence of SBI-0206965 in vitro homocysteine, we found genes encoding the serine

acetyl-transferase, CysE, the OAS-thiol-lyase, CysK, and two transporters CysP1 (Cpe0947) and CysP2 (Cpe0967) (Fig. 1 and 4). https://www.selleckchem.com/products/ferrostatin-1-fer-1.html T-box motifs are present upstream of cysK, cysP1 and cysP2 [42]. These T-box regulatory systems are mostly involved in the control of aminoacyl-tRNA synthetase genes but also of genes involved in amino-acid biosynthesis or uptake in firmicutes [11, 42, 43]. An alignment of the region surrounding the 15 bp T-box motif (AATTAGAGTGGAACC) located upstream of cysK, cysP1 and cysP2 is presented in Fig. 5. We clearly observed the presence of conserved AGTA-, AG-, GNUG- and F-boxes and a terminator downstream from the T-box motif with a possible alternative formation of an antiterminator structure. A specifier codon for cysteine (TGC) matching with the anticodon of the cysteinyl-tRNA is also present [11]. Interestingly, Vitreschak et al have shown that the TGC codon (100%) is preferred to the TGT codon at T-box regulatory sites [42]. The presence of a T-box specific for cysteine (T-boxCys) upstream of these genes is therefore in agreement with the 7 to 15.5-fold derepression under conditions of cysteine limitation in transcriptome. DNA Damage inhibitor This strongly suggests that these genes are controlled by premature termination of transcription

via a T-box element sensing cysteine availability. To confirm the control of cysP2 expression by premature termination of transcription, we performed Northern blot experiments using strand specific RNA probes located in the 5′ untranslated region of the cysP2 gene (T-box region) or within its coding region. In the presence of cystine, we observed small transcripts of about 500, 300 and 200 bases with a probe hybridizing with the T-box region while no transcript was detected with the cysP2 probe (Fig. 6). The transcript of 300 bases has the size expected for a transcript initiated at a putative σA-dependent promoter located upstream of the specifier hairpin (data not shown) and terminated at the T-box terminator (Fig. 5 and 6) while the band at 200 bases might be due to RNA degradation or cleavage of the 300 base transcript as observed for other T-box elements [44].