FIGURE 3.

Inhibition of 5S rRNA maturation by AS5. (A) Representation of the synthetic 5S-trnR transcript (top) and the complementary AS5-1 and AS5-2 transcripts aligned to the corresponding genome positions. (+1) First nucleotide of intergenic region; (P) RNase P cleavage site. (B, left two lanes) Labeled transcripts serving as markers for the migration of mature 5S rRNA (m5S) and the 5S-trnR precursor cleaved at the RNase P site (5S + I), respectively. (Third lane) 5S-trnR precursor; (fourth lane) precursor incubated for 30 min in buffer without chloroplast proteins. (Last 12 lanes) Uniformly labeled 5S-trnR transcript incubated with the chloroplast protein extract and increasing amounts of trace-labeled AS5 inhibitors. Products were separated in a 5% denaturing polyacrylamide gel. Icons at the left indicate the sense substrates, and arrows on the right indicate the antisense substrates.