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. Author manuscript; available in PMC: 2010 Sep 1.
Published in final edited form as: Mol Microbiol. 2009 Jul 21;73(5):882–897. doi: 10.1111/j.1365-2958.2009.06813.x

Figure 3. Deletion of RNases and A-site mRNA cleavage.

Figure 3

(A) Northern analysis of flag-(m)ybeL-PP transcripts isolated from RNase deletion strains. Deletions of genes encoding the various RNases were introduced into ΔtmRNA cells, and transcripts examined using the 5′ probe depicted in Fig. 1A. The migration positions of full-length, +18 truncated, and A-site cleaved transcripts are indicated. Samples marked with an asterisk (*) were taken from cells expressing FLAG-(m)YbeL-PP from a pACYC184-derived plasmid as described in Results. (B) Northern analysis of flag-(m)ybeL transcripts expressed in ΔRNase II cells. FLAG-(m)YbeL-PP (Pro-Pro) and FLAG-(m)YbeL-EA (Glu-Ala) proteins were expressed ΔtmRNA and tmRNA+ cells, with (RNase II+) or without RNase II (ΔRNase II). The positions of full-length and A-site cleaved transcripts are indicated. The assignment of +12, +18, and +28 truncation products in these samples was confirmed by S1 nuclease protection analysis (data not shown). (C) Northern analysis of A-site cleavage in ΔtmRNA cells lacking RNase III and RNase E activity. RNase E activity was assessed by shifting ΔtmRNA RNase E(ts) cells to the non-permissive temperature (42 °C). The migration positions of full-length, +18 truncated, and A-site cleaved transcripts are indicated.