Fig. 3.

Nus factors repress translation of suhB, leading to Rho-dependent termination within the gene. β-galactosidase activity of (a) transcriptional and (b) translational fusions of suhB to lacZ in wild-type cells, ΔnusB, nusE(A12E), or rho(R66S) mutants. The suhB-lacZ fusion had either a wild-type (‘wt’) or mutant boxA (‘C4T/T6C’). Data are normalized to levels in wild-type cells. Error bars represent ±1 standard deviation from the mean (n = 3). Schematics of constructs used in these experiments are depicted above the graphs. (c) and (d) Western blots showing SuhB-FLAG protein levels in wild-type cells, nusE(A12E), rho(R66S) c, and boxA(C4T/T6C) mutants d. SuhB-FLAG was probed with α-FLAG antibody; RNAP β′ was probed as a loading control. Representative blots from at least three independent experiments are shown. Images were cropped for clarity; unprocessed western blot images are in Supplementary Fig. 7