Observing the fate of tRNA and its modifications by nucleic acid isotope labeling mass spectrometry: NAIL-MS.

ABSTRACT: RNA in yeast, especially rRNA and tRNA are heavily modified to fulfill their function in protein translation. Using biosynthetic stable isotope labeled internal standards we quantified 12 modified nucleosides in tRNA from S. cerevisiae over 24 hours. We observed different quantities of modified nucleosides in dependence of the growth phase. To elucidate the underlying mechanism of the observed tRNA modification profile adaptation, it is necessary to distinguish the pre-existing tRNA pool and its modifications from newly-synthesized tRNAs. By combination of 2 differentially isotope labeled media we developed NAIL-MS, nucleic acid isotope labeling coupled mass spectrometry. During the yeast growth cycle we observe dilution of pre-existing tRNAs by newly-synthesized tRNAs by the growing numbe

SUBMITTER: Heiss M 

PROVIDER: S-EPMC5699550 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature