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. 2011 Jun 13;108(26):10502–10507. doi: 10.1073/pnas.1105645108

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Fig. 2. — Glucose starvation induces the accumulation of m¹G37-bearing tRNA^Phe in the nucleus. (A) LC/MS analyses of the total nucleosides in the tRNA^Phe molecules isolated from yeast cells (BY4742) after 0h (uppermost panels), 1h (second panels), 2h (third panels), and 3h (fourth panels) of glucose starvation. As a control, the nucleosides of the tRNA^Phe molecules that were isolated from ΔTYW1 are shown in the lowermost panels. The Left panels show the mass chromatograms of the base-related ion () of monomethylated guanosine (m/z 166), which detect m¹G and m²G. The Right panels show the mass chromatogram of the proton adduct of yWpA (m/z 838). (B) LC/MS RNA fragment analyses of the RNaseA-digested tRNA^Phe molecules. The panels show the mass chromatograms that detect the doubly charged ions of the anticodon-containing fragments that bear m¹G (Left panels, m/z 1,013.65) and yW (Right panels, m/z 1,119.20). The vertical axes display the apparent ratio of m¹G or yW, which is calculated on the basis of the intensity of the m¹G- and yW-containing fragments. (C) The TRM5-GFP (Upper) and TRM5-NES-GFP (Lower) constructs are shown. The photos show the phase contrast (PH), GFP-fluorescence, Hoechst 33342, and merged images for each strain. (D) LC/MS analyses of the total nucleosides in the tRNA^Phe molecules that were isolated from the TRM5-GFP (Upper) and TRM5-NES-GFP (Lower) strains after 4h of glucose starvation. The Left and Right panels show the mass chromatograms of the base-related ion () of monomethylated guanosine (m/z 166) and the proton adducts of yWpA (m/z 838), respectively.

Inline graphic — Glucose starvation induces the accumulation of m¹G37-bearing tRNA^Phe in the nucleus. (A) LC/MS analyses of the total nucleosides in the tRNA^Phe molecules isolated from yeast cells (BY4742) after 0h (uppermost panels), 1h (second panels), 2h (third panels), and 3h (fourth panels) of glucose starvation. As a control, the nucleosides of the tRNA^Phe molecules that were isolated from ΔTYW1 are shown in the lowermost panels. The Left panels show the mass chromatograms of the base-related ion () of monomethylated guanosine (m/z 166), which detect m¹G and m²G. The Right panels show the mass chromatogram of the proton adduct of yWpA (m/z 838). (B) LC/MS RNA fragment analyses of the RNaseA-digested tRNA^Phe molecules. The panels show the mass chromatograms that detect the doubly charged ions of the anticodon-containing fragments that bear m¹G (Left panels, m/z 1,013.65) and yW (Right panels, m/z 1,119.20). The vertical axes display the apparent ratio of m¹G or yW, which is calculated on the basis of the intensity of the m¹G- and yW-containing fragments. (C) The TRM5-GFP (Upper) and TRM5-NES-GFP (Lower) constructs are shown. The photos show the phase contrast (PH), GFP-fluorescence, Hoechst 33342, and merged images for each strain. (D) LC/MS analyses of the total nucleosides in the tRNA^Phe molecules that were isolated from the TRM5-GFP (Upper) and TRM5-NES-GFP (Lower) strains after 4h of glucose starvation. The Left and Right panels show the mass chromatograms of the base-related ion () of monomethylated guanosine (m/z 166) and the proton adducts of yWpA (m/z 838), respectively.