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Fig. 5 | BMC Biology

Fig. 5

From: Phosphoproteome dynamics mediate revival of bacterial spores

Fig. 5

Phospho-modifications of translational factors affect spore revival and vegetative growth. a Proteins were extracted from either spores or vegetative cells of the strains: (1) AR223 (Pxyl- rpsJ-HA, ∆rpsJ), (2) AR224 (Pxyl- rpsJ-S32A-HA, ∆rpsJ), and (3) AR225 (Pxyl-rpsJ-S32D-HA, ∆rpsJ). Equal amounts of protein extracts were subjected to Western blot analysis. Membrane was probed with antibody against HA-tag. b Strains PY79 (wild type, WT), AR168 (P xyl - rpsJ, ∆rpsJ), AR169 (P xyl - rpsJ-S32A, ∆rpsJ), and AR185 (P xyl -rpsJ-S32D, ∆rpsJ) were plated on LB supplemented with 0.5 % xylose and 7.5 μg/mL of tetracycline, incubated at 37 °C and photographed after 48 h. c Spores of PY79 (wild type, WT), AR168 (P xyl -rpsJ, ∆rpsJ), AR169 (P xyl -rpsJ-S32A, ∆rpsJ), and AR185 (P xyl -rpsJ-S32D, ∆rpsJ) strains were incubated at 37 °C in S7 defined medium supplemented with L-Ala (10 mM) + 0.5 % xylose, and optical density (OD600) was measured at the indicated time points. The data points are averages of results obtained from three independent biological repeats. Error bars designate SD. d Growth curves of PY79 (wild type, WT), AR168 (P xyl -rpsJ, ∆rpsJ), AR169 (P xyl -rpsJ-S32A, ∆rpsJ), and AR185 (P xyl -rpsJ-S32D, ∆rpsJ) strains. Cells were grown at 37 °C in S7-defined medium supplemented with L-Ala (10 mM) + 0.5 % xylose and OD600 was measured at the indicated time points. The data points are averages of results obtained from four independent biological repeats. Error bars designate SD. AR185 (P xyl -rpsJ-S32D, ∆rpsJ) showed significantly reduced growth rates compared to the other strains by repeated measures ANOVA (P <0.05). e Spores of AR168 (P xyl -rpsJ, ∆rpsJ) and AR185 (P xyl -rpsJ-S32D, ∆rpsJ) strains were incubated in S7-defined medium supplemented with L-Ala (10 mM) + 0.5 % xylose and monitored by time lapse microscopy. Shown are phase-contrast images acquired at the indicated time points. Arrows signify the dominating revival stages of each population at a given time point (germination, red; ripening, orange; elongation, green). Scale bar represents 1 μm

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