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. 2019 Sep 4;213(3):849–863. doi: 10.1534/genetics.119.302525

Figure 2.

Figure 2

Mutation of G1:C70 and C3:G70 reduces tRNASerUGG function allowing a nonlethal level of mistranslation. (A) Structures of four tRNASerUGG alleles created by flipping base pairs at the first four positions in the acceptor stem. (B) Wild-type strain (BY4742) expressing wild-type tRNASerUGA, tRNASerUGG-G26A or the viable tRNASerUGG-C1:G72 or -G3:C70 were grown to saturation in media lacking uracil and spotted in 10-fold serial dilutions on the same media. (C) Strains containing tti2-L187P (CY7020) and one of the tRNAs described in B were grown to saturation in media lacking uracil and spotted in 10-fold serial dilutions on media lacking uracil or complete media containing 5% ethanol. (D) Wild-type strain BY4742 containing one of the tRNAs described in B and a fluorescence heat shock reporter were grown to saturation in media lacking uracil and histidine. Cells were diluted 1:20 in the same media and grown for 6 hr. Cell densities were normalized and fluorescence measured. Each point represents one biological replicate.