Figure 4. Nucleoside analysis analysis in wild type Haloferax volcanii and the Δpcc1 mutant cells.

tRNAs were extracted from each strain and hydrolyzed to nucleosides. The nucleoside content was determined by HPLC with detection by UV/Vis at 254 nm. Analyses were performed in triplicate from independent cultures. A. HPLC chromatographs of nucleosides from wild-type, wild-type with synthesized t⁶A added, or Δpcc1. t⁶A elutes at 24 minutes. B. Comparison of the t⁶A peak area of wild-type and Δpcc1. The ratios of Ψ-modified base/m₂ ²G were used to normalize tRNA concentrations across samples with t⁶A peak area of wild-type set at 100%. Δpcc1 contains approximately 19% less t⁶A than wild-type (P = 0.02).