Fferences below the conditions tested (see Fig. S1 in the supplemental material). RNA-seq analysis on the regulon within the C. difficile R20291 agrA76a::CT mutant. Because the S. aureus agr locus plays a important role in controlling the coordinated expression of virulence genes, we used RNA-seq evaluation to start to define the regulon below the handle with the C. difficile R20291 agr locus. To this end, total RNA wasjb.asm.orgJournal of BacteriologyC. difficile agr Locusa)targeted intron 76|77a erm RAMC. difficile agr11 bases 61 bases225 aa438 aa46 aa193 aa (25 )agrALytTRREC(28 )agrC(23 )agrDagrBS. aureus agragrBagrDagrCb)C.difficile S.aureusMNRNFIYLLKIYKKGCLRIVISIGICDDELHYRIKIKDILSEILSSYPINYNIYEFSSGE 60 NRNFI LK KK CLR VISI RI IKD EI SY YNIYE SSGE -MKIFICEDDPKQRENMVTIIKN———YIMIEEKPMEIALATDNPYEVLEQAKN- 49 MK FI DD QR NMVTIIKN I IEEK MEI A YEVLE AKN : ** . :: : :*. * *:: ** : *:: * :.. LNN KDLD LIM M MDIQMKT INGM AR IREFD M MDT RK RE IIFVTSF VEFM YEV ELLNNYPKDLDILIMDIQMKT-INGMDTARKIREFDHKLEIIFVTSF–VEFMQEGYEVK 117 MN DIG YFLDIQLST INGIKLG EIRKHD L RK IIFVTSH LTYL VYKVA KV –MN—-DIGCYFLDIQLSTDINGIKLGSEIRKHDPVGNIIFVTSHSELTYLTFVYKVA 103 :* *:. ::***:.* ***:. . :**:.* :******. : :: *:* A YILK N KI K VL CIN EMMKKRNN TINVKN N NY VKN RIK DSI YI AYRYILKPINKEKISKSVLPCIN——EMMKKRNNYLTINVKN–YVDRIKIDSITYI 169 AM FIFK DD AELRTRIIDCLE LQLLSRDNSV TIELKR KR VQY DI FF YD AMDFIFK-DDPAELRTRIIDCLETAHTRLQLLSRDNSVETIELKRGSNSVYVQYDDIMFF 162 * :*:* : :: . :: *:: :::.: *. **::*. :: *.* :: ET TDR N IYTHDD KM SKIE NE FFRCHNSYIVN K VE MSGS VIV E ESM ETDRPNILIYTHDDMYITKMSISKIEKILNEYGFFRCHNSYIVNLKLVESMSGSTVIVDG 229 ES SS SHRL AHLD EFY N KE LD RFFRCHNSFV V VVN HNIE IDSK RIV E ES ESSTKSHRLIAHLDNRQIEFYGNLKELSQLDDRFFRCHNSFVVNRHNIESIDSKERIVYF 222 *:.5-Ethynyluridine Data Sheet .4-Bromo-2-methyl-1,3-thiazole structure : :* * :: .PMID:33480340 * : *******::** : :**:… ** KSIPISKYRVKGLKLAITNILGDIVC 255 KSI SK RVK KL K KGL KNKEHCYASVRNVKKI———- 238 KNK CY VR VKK K RNV *. . *:.:*10 three 4 5 6 7 8 9 2C.difficile S.aureusC.difficile S.aureusC.difficile S.aureusC.difficile S.aureusagrAREC signal receiver LytTR DNA bindingFIG 1 Genetic organization from the C. difficile 027 agr locus. (a) The complete R20291 agr locus (upper panel) in comparison with the agr locus of S. aureus. Arrows indicate genes and path of transcription. The length of every single protein is indicated above (aa, amino acids), plus the % identity towards the respective S. aureus orthologue is indicated below. The location of the group II intron target web page, in line with the ClosTron delivery system, applied to construct the R20291 agrA76a::CT mutant strain is shown. (b) ClustalW2 a number of sequence alignment of AgrA from C. difficile and S. aureus. Symbols: *, identical amino acids; :, strongly similar; ., weakly related. AgrA shares 28 amino acid identities, as determined by BLAST search. The two domains of AgrA are outlined: 5= REC signal receiver domain, green box; 3= LytTR-DNA binding domain, red box. The structure with the S. aureus AgrA LytTR-DNA binding domain (residues 137 to 238) is indicated (72). The secondary structure is indicated: helices, blue bars; strands, black bars.isolated from C. difficile R20291 and R20291 agrA76a::CT grown to late exponential phase. The log2 fold alter of transcript abundance amongst these RNA populations was obtained (P 0.1) (see Fig. S2 in the supplemental material), and transcripts had been visualized by mapping the coverage per base pair towards the reference R20291 genome (Fig. 2a). In total, 75 transcripts have been discovered to be drastically differentially expressed in th.
