Actin-binding protein ABP140 is a methyltransferase for 3-methylcytidine at position 32 of tRNAs in Saccharomyces cerevisiae

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FIGURE 2.
FIGURE 2.

Mass spectrometric analysis of total nucleosides and individual tRNAs from S. cerevisiae wild-type and mutant cells. (A) LC/MS analysis of total nucleosides from strains of wild-type (WT), yor239wΔ, yor240wΔ, and abp140Δ. Graphs show mass chromatograms traced by MH+ (m/z 258) of m3C, m5C, and 2′-O-methylcytidine (Cm). Arrows indicate the retention time for m3C. (B,C) CapLC/nanoESI MS analysis of RNase T1-digested tRNAThr1 (B) or tRNASer1 (C) isolated from wild-type (top) and yor240wΔ (bottom) strains. (Left) The mass chromatograms traced by doubly charged ions of fragments bearing m3C32 (m/z 1602 [B] or 1679 [C]) and C32 (m/z 1595 [B] or 1665 [C]). (Right) The mass spectra of position 32 containing fragments. The charge states of multiply charged ions are indicated in parentheses. The RNA sequences of each fragment are indicated on each graph. “>p” at the 3′ end of the fragment indicates cyclic phosphate. “Relative abundance” for y-axis of mass chromatograms and mass spectra shown in this figure and Figures 4, 5, and 6 stands for the percentile presentation of peak intensities that were normalized, with the highest peak defined as 100.

This Article

  1. Published in Advance April 25, 2011, doi: 10.1261/rna.2653411 RNA 17: 1111-1119

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