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. 2022 Sep 9;8(36):eabp8655. doi: 10.1126/sciadv.abp8655

Fig. 4. Mutation near template exit channel increases RNA polymerase sensitivity to t-loops.

Fig. 4.

(A) Structure of the pretermination complex of the bat IVA RNA polymerase (PDB 6SZU). The 5′ and 3′ ends of the template are shown in black and gold, respectively. The body of the template is shown in dark blue, and the nascent RNA is shown in red. Location of PB1 K669 and PB2 T81 is indicated. (B) Amino acid alignment of PB1 C terminus. PB1 K669 is indicated with an arrow. (C) IFN-β promoter activity and segment 6 vRNA levels in the presence of WT and K669A RNA polymerases. For the segment 6 vRNA template, the mRNA, cRNA, and vRNA species are indicated. 5S rRNA and Western blot for PB1 subunit expression are shown as loading control. (D and E) IFN-β promoter activity and mvRNA template levels for five NP71 mvRNA templates in the presence of WT and K669A RNA polymerases. RNA levels were analyzed by primer extension. 5S rRNA and Western blot for PB1 subunit, NP, and tubulin expression are shown as loading control. Data from three biological repeats are shown.