Figure 4 | Scientific Reports

Figure 4

From: Accurate target identification for Mycobacterium tuberculosis endoribonuclease toxins requires expression in their native host

Figure 4

tRNA modifications are not required for VapC-mt11 target recognition in vitro. Total M. tuberculosis RNA was incubated with (+) or without (−) recombinant VapC-mt11. Northern analysis of (a) tRNAArg27 (b) tRNAArg40 (c) tRNAGln32 (d) tRNAGln41 (e) tRNAPro14 (f) tRNAPro23 (g) tRNAPro35 (h) tRNALeu3 (i) tRNALeu13. The light band in the + lane of panel b is background, not uncleaved tRNA. Position of the oligonucleotides used are shown by the red dots on the tRNA diagrams above each bracketed group. Cleavage positions, yellow arrow. Oligonucleotides were designed to hybridize to the ASL to optimally differentiate between tRNA species. Therefore, cleavage products were generally not visible because the oligonucleotides could no longer hybridize to the cleaved ASL. Hybridization temperatures were optimized to preclude cross-hybridization to other tRNAs. tRNA numbering and anticodon sequences of each numbered tRNA from the Lowe lab genomic tRNA database http://gtrnadb.ucsc.edu33. All uncropped images shown in Supplementary Information Fig. 3.

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