Substrate recognition and auto-inhibition in the central ribonuclease RNase E. Bandyra et al.
Description
Images of electrophoretic gels collected under denaturing conditions for RNA degradation assays recorded with GeneSnap from Syngene. The gels were stained with SybrGold and visualised by uv fluorescence. Figure 1D shows 9S processing by RNase E 1-529 wild type and mutants. Figure 1E shows ompD degradation in the presence of 12-mer MicC for RNase E 1-529 wild type and mutants. Figure 2D shows 9S processing by RNase E 1-529 wild type and super-activating triple mutant. Figure 3a, b and c show ompD 50-mer processing in the absence and presence of 12-mer MicC by RNase E 1-529 wild type. Figure 5B shows 9S processing by RNase E 1-529 wild type and 8x mutant on the duplex recognition surface. Figure 5C shows artificial substrate 62-mer with 2 stem loops degradation by RNase E 1-529 wild type and 8x mutant on the duplex recognition surface. Supplementary Figure 3A shows 9S processing by full degradosome and truncated degradosome (RNase E 1-850, helicase and enolase). Supplementary Figure 3B shows degradation of the artificial 62-mer with the 2 stem loops by truncated degradosome (RNase E 1-850, helicase and enolase). Supplementary Figure 3C shows degradation of the artificial 62-mer with the 2 stem loops by full degradosome.
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Steps to reproduce
Procedure described in Bandyra et al. (2018) Substrate recognition and auto-inhibition in the central ribonuclease RNase E. Molecular Cell (in press).