Figure 3.

Ψ at position 2819 in yeast mitochondrial 21S rRNA disappears upon deletion of the YLR165c ORF. Primer extension analysis of the CMCT-modified S.cerevisiae mitochondrial 21S rRNA in a total RNA fraction from wild-type (WT) or ΔYLR165c S.cerevisiae strains, for 1 (lanes 2 and 8), 10 (lanes 3, 5, 9, 11) and 20 min (lanes 4, 6, 10, 12). A control extension experiment was made without addition of CMCT (lanes 1 and 7). The experimental conditions are the same as in Figure 2. The reverse transcription stop, corresponding to residue Ψ₂₈₁₉, is indicated by an arrow on the left. Absence of this stop in the reverse transcription pattern of the 21S rRNA from the ΔYLR165c strain (lanes 5 and 6) indicates the absence of residue Ψ₂₈₁₉. Primer extension analysis of the CMCT-modified S.cerevisiae mitochondrial 21S rRNA in a total RNA fraction from the ΔYLR165c strain transformed with plasmid p413TEF-YLR165c (lanes 9 and 10) and with a plasmid bearing mutated YLR165c ORF (lanes 11 and 12).