FIG 5.

Analysis of chain termination effects of ribonucleotide analogs after incorporation into RNA synthesized by POLRMT. (A) Chemical structures of the ribonucleotide 5′-triphosphate analogs tested in the assay. (B) The primer and template used in the assay whose results are shown here. (C) Analysis of the incorporation and chain termination abilities of the ribonucleotide analogs. Primer extension reactions were initiated by the addition of ribonucleotide analogs at 100 μM and of three complementing natural ribonucleotides at 1 μM each, as indicated under each lane. The reactions were performed at 22°C for 1 h. The products of the primer extension reaction were resolved by denaturing PAGE. The natural ribonucleotides to be incorporated are shown on the sides of the graph. The natural rNTP concentrations in lanes 1, 2, and 3 are 100 μM, 10 μM, and 1 μM, respectively.