Figure 6. Rhomboid cleavage licenses further degradation of substrates.

• A
  Western blot analysis (probing with an anti‐V5 mAb) to detect degradation of N‐terminally V5‐tagged wild‐type (WT) or modified (P³⁰⁰A) HybA in S. sonnei Δrhom7 chromosomally expressing wild‐type (+) or inactive (S²⁰¹A) GlpG with (+) or without (−) HybB.
• B
  Degradation of V5‐tagged HybA at times after blocking protein translation at T₀ in the presence (+) or absence (−) of HybB.
• C
  Western blot analysis of N‐terminally V5‐tagged wild‐type (WT) or modified (P²⁵⁹A) FdoH in S. sonnei Δrhom7 chromosomally expressing wild‐type (+) or inactive (S²⁰¹A) GlpG with (+)/without (−) FdoI.
• D
  Western blot analysis of N‐terminally V5‐tagged wild‐type (WT) or modified (P²⁵⁹A) FdnH in S. sonnei Δrhom7 with wild‐type (+) or inactive (SAHA) Rhom7 expressed from pBAD33 with (+)/without (−) FdnI.
• E, F
  Degradation of N‐terminally V5‐tagged wild‐type (WT) or modified (P²⁵⁹A) FdoH (E) or FdnH (F) in S. sonnei Δrhom7 with wild‐type (+) or inactive (S²⁰¹A, SAHA) GlpG expressed chromosomally (native) or from pUC19 (plasmid) without FdoI or FdnI (−), respectively, +/− exposure to 400 μM CuCl₂ for 30 min.

Data information: Rhomboid substrates that are uncleaved, cleaved by GlpG or cleaved by Rhom7 are marked by black, red and blue arrows, respectively. Degradation products post‐rhomboid cleavage are marked by green arrows. RecA, loading control.Source data are available online for this figure.