FIG 5.

Rip1 is required for the proteolytic processing of RsdA. (A) Experimental schematic for degradation of RsdA with an N-terminal MBP tag in the mycobacterial membrane. Shown are the possible species of MBP-RsdA after proteolysis by site-1 (S1P) and site-2 (i.e., Rip1) proteases. Lysates from the M. smegmatis wild type (WT) and ΔMSMEG_2579 mutant (ΔMsrip1) with plasmids encoding MBP-RsmA (B), MBP-RsdA(C), and MBP-RsgA (D) were analyzed by immunoblotting with antibodies recognizing MBP or RNAPβ as a loading control. (E) Immunoblots for MBP (top) or RNAPβ (bottom) of MBP-RsdA expressed in biologic triplicates of WT or Δrip1 M. tuberculosis.