Investigation of the activity of tRNA modification enzymes Trm732 and Trm734

Abstract

Transfer RNAs (tRNAs) are molecules that transport amino acids to the ribosome to facilitate protein synthesis. Post-transcriptional modifications of tRNA are required for function. Trm7 is a methyltransferase enzyme which interacts with Trm732 and Trm734 to modify tRNAs at residues C32 and G34, respectively. Trm732 and Trm734 variants have been generated via site-directed mutagenesis and transformed into Saccharomyces cerevisiae. Analysis of these variants, when compared to wild type, allows for the identification of the residues involved in the methylation of tRNA. Function has been investigated quantitatively by measuring nucleoside modification levels with Ultra-Pressure Liquid Chromatography, and qualitatively through growth assays.