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. 2015 Jan;21(1):61–74. doi: 10.1261/rna.047639.114

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© 2014 Guy and Phizicky; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 3. — The biologically important Sp Trm7 tRNA substrate is tRNA^Phe. (A) Overexpression of tRNA^Phe suppresses the near lethality of S. pombe trm7▵ mutants. Strains with plasmids as indicated were grown overnight in EMM-Leu and analyzed as in Figure 2A, after incubation for 3 d at 30°C. (B) Overexpression of tRNA^Phe in S. pombe trm7▵ mutants does not completely restore wild-type growth at 37°C. Strains as indicated were purified on 5-FOA to ensure loss of the [P_nmt1** Sp trm7⁺ ura4⁺] plasmid, grown overnight in EMM-Leu, and then growth was analyzed as indicated in Figure 2A. (C) tRNA^Phe from the S. pombe trm7▵ [tRNA^Phe] strain lacks yW. tRNA^Phe purified from the indicated strains after growth in EMM-Leu was analyzed as described in Figure 2C.