Chapter 11. Identification and analysis of tRNAs that are degraded in Saccharomyces cerevisiae due to lack of modifications.
Mounting evidence shows that tRNA modifications play crucial roles in the maintenance of wild-type levels of several tRNA species. This chapter describes a generalized framework in which to study tRNA turnover in the yeast Saccharomyces cerevisiae as a consequence of a defect in tRNA modification status. It describes several approaches for the identification of tRNA species that are reduced as a consequence of a modification defect, methods for analysis of the rate of tRNA loss and analysis of its aminoacylation, and methods for initial characterization of tRNA turnover. These approaches have been used successfully for several modification defects that result in tRNA turnover.
Chernyakov
Irina
I
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York, USA.
Baker
Melanie A
MA
Grayhack
Elizabeth J
EJ
Phizicky
Eric M
EM
D015152:Blotting, Northern; D020411:Oligonucleotide Array Sequence Analysis; D020871:RNA Stability / Q000235:genetics*; D012343:RNA, Transfer / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics
D012343:RNA, Transfer