RESEARCH 2003
RESEARCH 2002
RESEARCH 2001
RESEARCH 2000

Tracy Ross
Alcorn State
Summer Mentor: Dr. James Anderson

The tRNA M¹ A⁵⁸ methyltransferase is composed of two subunits encoded by the genes TRM6 and TRM61. Mutations in TRM6 lead to reduced levels of initiator tRNA compared to the wild type. The reduced levels of initiator tRNA in a trm6-504 temperature sensitive mutant is due to its degradation. Genetic studies have shown that TRF4, a DNA and Poly A polymerase and the exosome are involved in the degradation of aberrant initiator tRNA. Whereas, deletion of TRF4 leads to stabilization of tRNAi^Met, overexpression of Trf4p destabilizes the hypo-modified tRNAi^Met in trm6-504. The question to be address in my project is whether or not tRNA surveillance and degradation is limited to a discrete region within the nucleus. A Green Fluorescent Protein (GFP) was fused to the TRF4 gene in TRM6 WT and trm6-504 mutant strain to determine localization of TRF4 in WT and mutant yeast. TRF4:GFP fusion protein localized to the nucleus in WT cells. However, fusion of GFP to TRF4 in trm6-504 mutant has not succeeded. Methods involved in this study include PCR Amplification, Transformation, and Isolation of Genomic DNA from Yeast. Observation of the changes or similarities was performed using UV microscopy.