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. 2008 Mar 10;28(10):3301–3312. doi: 10.1128/MCB.01542-07

FIG. 2.

FIG. 2.

Hypomodification induced by elp3Δ, tuc1Δ, or trm9Δ alleles does not reduce abundance or aminoacylation of any of the affected tRNA species. Northern blot analyses of total RNA isolated under either basic (A) or acidic (B) conditions from wild-type (W303-1B), elp3Δ (UMY2843), tuc1Δ (UMY3165), or trm9Δ (UMY3297) cells. After polyacrylamide gel electrophoresis and transfer under appropriate conditions, blots were probed for the U34-containing tRNA of interest using 32P-labeled oligonucleotides. To control for loading and indirect effects on aminoacylation, the blots were also probed for the C34-containing initiator methionine tRNA (tRNAiMet). (A) Steady-state tRNA levels. (B) Aminoacylation levels. The first lane in each individual blot in panel B represents a deacylated wild-type RNA sample. The symbols for the U34-containing tRNAs indicate the anticodon sequence of wild-type cells. The elp3Δ strain lacks the mcm5 and ncm5 side chains whereas the tuc1Δ strain lacks the s2 groups. Cells deleted of TRM9 lack the esterified methyl group of mcm5 side chains. wt, wild type.