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. 2007 May 16;81(15):7974–7984. doi: 10.1128/JVI.00714-07

FIG. 3.

FIG. 3.

Mutations in stem-loop I but not stem-loop II of U1 RNA abolished inhibition of polyadenylation of P41-generated RNAs at (pA)p. (A) RPA, using the DHA2CAA probe, of RNA isolated from 293 cells transfected with reporter plasmid P41CapA2CAADm, together with either the control plasmid SK or various U1S and U1S mutants as described in Materials and Methods and as indicated on the top of each lane. Bands representing RNAs that either read through or are polyadenylated at (pA)p are designated (pA)d or (pA)p, respectively. Quantifications of the ratios of P7-generated RNA at (pA)p to total P7-generated RNA are shown as percent (pA)p and are averages of the results for at least three individual experiments, with standard deviations underneath. (B) Diagram of the putative interaction of the U1S snRNA with the P41CapA2CAADm reporter plasmid. The curved arrow indicates that the U1 snRNP has an inhibitory effect on polyadenylation at (pA)p. (C) The individual mutations for stem-loop I mutants U1SΔ70K, U1SΔ70Kb, U1SΔ70Kg, and U1SLoopI→II; stem-loop II mutants U1SΔA, U1SΔAg, and U1SLoopII→I; and stem-loop I and II mutant U1SΔ70K+ΔA are indicated.