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. 2012 May 7;109(21):8050–8055. doi: 10.1073/pnas.1205990109

Fig. 2.

Fig. 2.

A complex of T7 gp5.5, E. coli H-NS, and tRNA. (A) Gp5.5 copurified with E. coli H-NS and its expression is eliminated by suppressor mutations. T7 gene 5.5, gene 5.5/5.7, and gene 5.5/5.7 from suppressors 11–51 were overexpressed as His-tagged proteins and purified by Ni-NTA before loading onto the SDS/PAGE gel. (B) Gp5.5 is copurified with RNA. Purified gp5.5/H-NS (1 μM) from cells expressing gp5.5 was electrophoresed through an agarose gel and then stained with ethidium bromide to detect nucleic acids. The gp5.5/H-NS complex was incubated with DNase I or RNase I (1 U/μL) for 1 h at 37 °C before loading. (C) Mobility of RNA components in gp5.5/H-NS complex is similar to E. coli tRNAs. The purified complex, RNA components extracted from the complex, and E. coli total tRNAs were compared on a 15% TBE-Urea gel. (D and E) Comparison of the RNA components in gp5.5/H-NS complex and E. coli tRNAs for their ability to be charged with leucine (D) and arginine (E) using E. coli aminoacyl-tRNA synthetases.