Skip to main content
. Author manuscript; available in PMC: 2017 Oct 11.
Published in final edited form as: Biochemistry. 2016 Sep 30;55(40):5649–5652. doi: 10.1021/acs.biochem.6b00920

Figure 3.

Figure 3

FtsH degrades ProW1-182-FLAG-ssrA. (A) Pulse-chase experiments demonstrate that ProW1-182-FLAG-ssrA has a longer half-life in cells lacking FtsH compared to wild-type cells or those lacking ClpP, ClpX, or ClpA (see Fig. 1B legend for experimental details). (B) Auto-radiograms of whole-cell lysates from pulse-chase experiments show that endogenously ssrA-tagged ProW1-182-FLAG-trpAt is also stabilized in a strain lacking FtsH. (C) Transient induction of plasmid-encoded ProW1-182-FLAG-ssrA in E. coli AR3289 ΔftsH::Kan resulted in little degradation (top panels) unless expression of plasmid-encoded Myc-tagged FtsH was subsequently induced (bottom panels). In all panels, ProW1-182-FLAG-ssrA expression was induced with IPTG for 10 min, and then pGAL was added to repress further expression. In the bottom panels, expression of Myc-tagged FtsH was induced by addition of ATc two min after addition of pGAL. Samples were separated by SDS-PAGE and immunoblotted with anti-FLAG and anti-Myc tag antibodies.