Base-pairing and stacking in the H1–H94–H98 nucleotide quartet. (A) Overview of the nucleotide quartet in the WT 70S ribosome. Hydrogen bonds between the bases and sugars are shown in black. (B) nLuc in vitro translation assay for WT (no tag), MS2-V2 inserted into H98, and MS2-V2 inserted into H25 after preincubation of 50S subunits at 60°C. Data are normalized to untagged 50S subunits. (C) nLuc in vitro translation assay for 50S subunits with MS2-V2 inserted into H25 and mutations in the H1–H94–H98 quartet after preincubation of 50S subunits at 60°C. Data are normalized to subunits with MS2-V2 inserted into H25. All error bars are the standard deviation of three experimental replicates.
