FIG 8.

YbeY controls the translation of RpoS through ReaL. (A) Wild-type PA14, the ΔybeY mutant, and the complemented strain were grown in LB to an OD₆₀₀ of 1. The relative levels of the sRNAs reaL and rgsA were determined by real-time PCR. Results represent means ± SD. ***, P < 0.001 by Student’s t test. (B) PA14, the ΔybeY mutant, and the ΔybeY ΔreaL mutant containing the RpoS(F1)-FLAG translational fusion were cultured in LB to an OD₆₀₀ of 1 and then incubated with 0.2% l-arabinose for 60 min. The bacteria were collected for Western blot analysis. The relative mRNA levels of the rpoS-Flag were determined by real-time PCR using the primers matching to the Flag tag RNA. The results represent means ± SD. ns, not significant. (C) The potential base-pairing between ReaL and the 5′ UTR is indicated by asterisks. The mutated nucleotides are underlined. PA14 and the ΔybeY mutant containing the indicated rpoS-Flag fusions were cultured in LB to an OD₆₀₀ of 1 and then incubated with 0.2% l-arabinose for 60 min. The bacteria were collected for Western blot analysis. (D and E) The indicated strains were grown in LB to an OD₆₀₀ of 1. The relative mRNA levels of katA (D) and rpoS (E) were determined by real-time PCR. The results represent means ± SD. **, P < 0.01 by Student’s t test; ***, P < 0.001 by Student’s t test.