Fig 8. Loss of function of AtTRM5 affects tRNA halve abundance.
(A) Northern blot analysis of RNA from wild type, trm5-1 and 35S:TRM5 trm5-1. tRNA-Ala (AGC) was inspected for changes in m1G37 and tRNA-Asp (GTC) and m1I37. tRNA-Arg (CCT) and rRNA were used as loading controls. The blots were probed with either tRNA-Arg (CCT), tRNA-Asp (GTC), or tRNA-Ala (AGC). (B) Accumulation and aminoacylation of tRNA-Ala (AGC) in wild-type plants, trm5-1 mutant plants and 35S:TRM5 trm5-1. The aminoacylated tRNA migrates slower than its corresponding deacylated species. To visualize the difference in electrophoretic mobility, aliquots of the same samples were deacylated in vitro. 10 μg of RNA was separated by electrophoresis, blotted and hybridized to a tRNA-Ala (AGC)-specific probe. The 25S rRNA band of the ethidium bromide-stained gel prior to blotting is shown as a loading control.