Protein Complexes and Proteolytic Activation of the Cell Wall Hydrolase RipA Regulate Septal Resolution in Mycobacteria (original) (raw)
Figure 7
Processed RipA is toxic to M. tuberculosis.
(A) Full length RipATB and truncated RipATB-AD was overexpressed in M. tuberculosis for several days with addition of aTc. Cells were analyzed by microscopy for changes in morphology. Membranes were stained with FM4-64. Scale bar represents 2 µM. (B) Growth of M. tuberculosis induced with aTc and overexpressing full length RipATB was measured by OD600. (C) Growth of M. tuberculosis induced with aTc and overexpressing truncated RipATB-AD was measured by OD600. (D) M. tuberculosis RipA (RipATB) was overexpressed in M. tuberculosis by induction (lane 2) with aTc for 48 hours. RipATB was then detected by anti-RipA Western blot analysis. Uninduced lysates were used a control (lane 1). Full length RipA (arrow) and processed forms (brackets) were detected. (E) Total overexpressed protein was quantified from (D) by performing densitometry analysis on bands apparent in the induced strain that are absent from the uninduced control. Fold change in total RipATB overexpression relative to endogenous full length RipA signal was graphed.