Figure 2
From: A novel regulator CdsR negatively regulates cell motility in Bacillus thuringiensis
Transcription of hag1, hag2, yjbJ-flgG, and yaaR-fliG2 operons/genes are regulated by CdsR. (A) RNA-seq data were analysed using Rockhopper software (version 2.03) and operons in the che/fla gene cluster were predicted in B. thuringiensis. Genes of the same color in the figure belong to a single operon. Open reading frames (ORFs) are represented by broad arrows, and promoter regions are represented by black arrows. The different operons in the figure are described in Table 1. The genetic organization of the fla gene cluster, including the yaaR-fliG2, hag1, hag2, and yjbJ-flgG operons, are represented by green, dark red, light red, and blue arrows, respectively. The numbers at the ends of the lines represent the locations in the genome. (B) Schematic representation of the B. thuringiensis flagellum. The flagellum is divided into three structural parts: the helical filament, which acts as a propeller; the basal body, which is embedded in the cytoplasmic membrane; and the hook, which connects the filament to the basal body. The basal body acts as a bidirectional rotary motor consisting of a rotor ring complex and several stator units around the rotor. (C) β-galactosidase activity of Phag1-lacZ in the cdsR mutant strains and HD73 wild-type strain. T0 is the end of exponential phase, and Tn is n hours after T0. Each value represents the mean of at least three replicates. (D) EMSA experiment confirms CdsR binding to hag1 gene promoter fragment (317 bp). The last lane, incubation of 200-fold greater unlabeled Phag1 probe mixed with the labeled Phag1 probe and 1.79 µM CdsR. (E) β-galactosidase activity of Phag2-lacZ in ΔcdsR and HD73. (F) EMSA experiment confirms CdsR binding to hag2 gene promoter fragment (610 bp). (G) β-galactosidase activity of PyjbJ-flgG-lacZ in ΔcdsR and HD73. (H) EMSA experiment confirms CdsR binding to yjbJ-flgG operon promoter fragment (316 bp). (I) β-galactosidase activity of PyaaR-fliG2-lacZ in ΔcdsR and HD73. (J) EMSA experiment confirms CdsR binding to yaaR-fliG2 operon promoter fragment (235 bp).
