Full Text
An Adenosine Deaminase that Generates Inosine at the Wobble Position of tRNAs
André P. Gerber and Walter Keller

Supplementary Material

Supplementary Figure 1.

Modifications in yeast TrnaAla. Adenosine (A) at position 34 (wobble position) is converted to inosine (I) by the tRNA-specific adenosine deaminase Tad2p/3p. A at position 37 is deaminated by Tad1p and further methylated to N1-methylinosine (see text for references).

Medium version | Full size version

Supplementary Figure 2.

Tad2p and Tad3p form a heterodimer. 40 μg of rTad2p and 25 �g of rTad3p were combined and passed over a 125 ml SuperdexTM 200 gel filtration column (HiLoadTM 16/60 prep grade, Pharmacia). One ml fractions were collected and 2 μl were used in a tRNA-specific deamination assay. The activity peak eluted at 73 ml. Arrowheads indicate the positions of marker proteins with their molecular masses (ferritin, 444 kD; aldolase, 158 kD; albumin, 67 kD; ovalbumin, 43 kD; RNase A, 13.7 kD).

Medium version | Full size version

Supplementary Figure 3.

The sequences of Tad2p and Tad3p are related. Multiple sequence alignment of Tad3p, Tad2p and putative homologous proteins. Conserved residues (>70%) are depicted within a black frame and similar amino acids are boxed in grey. Two conserved motifs (overlined) containing putative Zn2+-chelating residues (#) and a proton-transferring amino acid (o) are indicative for the deaminase domain. The positions of spliced introns in Tad3p are marked by arrows, a putative nuclear localization (NLS) is framed and peptide fragments confirmed by mass-spectrometry analysis are underlined.

Medium version | Full size version