Extended Data Fig. 1: In S. aureus, SprF1 inhibits global translation in vivo but not transcription.
The S. aureus strains N315_pCN35 (WT), N315ΔsprG1/ΔsprF1_pCN35 (ΔΔ), N315ΔsprG1/ΔsprF1_pCN35ΩsprF1 (ΔΔ + SprF1), N315ΔsprG1/ΔsprF1_pCN35ΩsprG1/sprF1 (ΔΔ + SprF1 + SprG1) were cultivated in diluted TSB medium until the exponential growth phase. a, After RNA extraction, northern blot analysis was done on SprG1 and SprF1 expression, with 5 S rRNA used as the loading control (n = 3). b, Growth kinetics of the four S. aureus strains. Error bars show the means and standard deviations of three independent experiments (n = 3). c, The S. aureus strain N315_pCN35 (WT) was cultivated 1 h at 37 °C and incubated with [3H]-uridine in the absence or presence of 20 µg/mL rifampicin or streptomycin over 1 h at 37 °C. The incorporation of the [3H]-uridine into the transcriptome was determined after TCA precipitation and filtration of S. aureus lysates. The incorporation was quantified and is shown compared to the radiolabelled RNAs of the untreated cells (Control), arbitrarily set to 1 (n = 3). d, The strains were incubated for 3 h at 37 °C with [35S]-methionine, with 10 µg/mL chloramphenicol (CHL) used as the positive control. Shown are the results of a representative SDS-PAGE of the newly synthesized radiolabeled proteins, with Coomassie blue staining as the loading control (n = 3). Graph bars represent the mean ± SD and black dots represent individual data points for three independent experiments. Statistical significance was calculated with a two-tailed unpaired t-test and p value are indicated: ***, p < 0.001. Source data are provided as a Source Data file.
