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. 2019 Jun 21;8:e46313. doi: 10.7554/eLife.46313

Figure 5. Orn acts as a diribonuclease in cell lysates.

Degradation of 32P-GG (A) and 32P-AAAAAGG (B) by whole cell lysates of wild-type, orn mutant, orn mutant complemented with ornVc, or ornVc D12A. For ∆orn and ∆orn complemented with ornVc D12A,100 nM of purified OrnVc was added at 30 min time point and incubated for an additional 10 min. Samples were stopped at the indicated time and analyzed by 20% denaturing PAGE. Representative gel images of triplicated assays are shown with the indicated RNA size. Graphs show quantitation of triplicate data for indicated RNA species over time.

Figure 5—source data 1. Numerical data for Figure 5.

Figure 5.

Figure 5—figure supplement 1. Catalytic residues of OrnVc are required for degradation for 32P-AAAAAGG in whole-cell lysates.

Figure 5—figure supplement 1.

Degradation of 32P-AAAAAGG by whole-cell lysates of the indicated strain (PA14, ∆orn mutant complemented with pMMB-ornVc L18A or pMMB-ornVc H158A. For ∆orn expressing ornVc L18A or ornVc H158A, 100 nM of purified, wild-type OrnVc protein was added at the 30 min time point and reactions were stopped 10 min thereafter. Samples were stopped at indicated time and analyzed by denaturing 20% PAGE. Representative gel images of two independent experiments are shown.