Molecular identification and phenotypic characterization of let-65 (mars-1) in Caenorhabditis elegans.

let-65 was first identified in a screen for lethal mutations in C. elegans’unc-22 region. I have demonstrated the molecular identity of let-65 to be F58B3.5 (mars-1), which encodes a methionyl tRNA synthetase (MARS-1). To develop a deeper insight into MARS-1 activity, I experimentally confirmed, from its sub-cellular localization, that it is a cytoplasmic enzyme. I also attempted to determine the sub-cellular localization of every known C. elegans amino-acyl tRNA synthetase using computational methods and, in collaboration with WormBase, renamed the genes appropriately. In addition, I studied let-65 transcription regulation by analyzing its 5’ promoter containing region. Haplo-insufficiency phenotypes manifest as a consequence of reduction in copy number of genes that encode proteins involved in translation. I have investigated this in C. elegans. I used two alleles of each of let-65 (mars-1) and let-336 (rps-27) and examined putative haplo-insufficiency phenotypes for both these genes.