Figure 7
In vivo validation of potential sRNA::mRNA interactions between teg58 and argGH transcript and mRNA stability in S. aureus cells. (A) Growth analysis in CDM containing 1% glucose without supplemented l-arginine for various isogenic strains as indicated. These strains include the wild type JE2, isogenic teg58 mutant; teg58 mutant complemented with native teg58, argG::ermC; argH::ermC; teg58 mutant with shuttle vector pSK236 carrying teg58 with the deletion of 8-nt interaction site for argG; teg58 mutant with shuttle vector pSK236 carrying teg58 with the 9-nt interaction site for argH scrambled; chromosomal mutation of the argG interaction site in teg58 (9-nt or 3 codons/AFA deletion); chromosomal mutation of the argH interaction site in teg58 (9-nt scrambled or SLQ to SLT). (B) Biofilm formation for various strains as indicated and described in (A) in TSB with added 0.25% glucose for 24 h in a microtiter plate. (C) argG mRNA stability in various strains was evaluated by qRT-PCR (2−ΔΔCT) using total RNA isolated from respective strains at the exponential phase of growth after treatment with rifampicin (200 μg per mL) at different time points. The data were normalized against rpoB as the reference transcript for the qRT-PCR. Relative expression of the gene (argG) in the wild type or teg58 mutant was plotted against no rifampicin treatment set as 100%. Means and standard deviations from triplicate experiments are shown. Student t-test (**P < 0.01; *** or ****P < 0.001).
