Poster abstracts
Poster number 44 submitted by Alisha Detmer
Investigating the Interaction Between Yeast tRNA Modification Proteins Trm7 and Trm734
Alisha Detmer (Chemistry and Biochemistry, Northern Kentucky University ), Julia Verhoff (Chemistry and Biochemistry, Northern Kentucky University), Holly Funk (Chemistry and Biochemistry, Northern Kentucky University), Michael P. Guy (Chemistry and Biochemistry, Northern Kentucky University)
Abstract:
In the anticodon loop of tRNAPhe, the modification of nucleotide G34 is fundamental for translation in Saccharomyces cerevisiae, humans, and other eukaryotes. In S. cerevisiae, the 2'-O-methylation of G34 is done by the Trm7:Trm734 complex. We study this complex because it is not fully understood why Trm7 needs Trm734 to function. The TRM7 human homolog is FTSJ1, and the TRM734 homolog is WDR6. Loss of function mutations in FTSJ1 that cause loss of Gm34 modification in patients result in non-syndromic x-linked intellectual disability. Through sequence alignment of Trm7 homologs in a variety of species and analysis of the crystal structure of the yeast Trm7:734 complex, Trm7 variants were made by changing amino acids which were conserved in different species, or that had a salt bridge, hydrogen bond, or high buried surface area with Trm734. A growth test that forces Trm7 to interact with only Trm734 or only its other binding partner Trm732 in yeast cells showed that some of the chosen amino acids were important for the Trm7:Trm734 interaction, but not for the Trm7:Trm732 interaction. Subsequent immunoprecipitation experiments further showed that these Trm7 residues were important only for Trm734 interactions, although addition of a tag to Trm7 appeared to result in some interference with Trm734 binding.
Keywords: tRNA , methylation, yeast