Post-transcriptional modification is an important part of translation regulation. FTSJ1 is a methyltransferase that interacts with binding proteins to perform post-transcriptional 2'-O-methylation on tRNAPhe. In humans, mutated FTSJ1 is associated with non-syndromic X-linked intellectual disability. In yeast, Trm7, Trm734, and Trm732 are the orthologs of FTSJ1, WDR6, and THADA, respectively. Trm7 performs 2'-O-methylation on tRNAPhe by separately interacting with Trm732 and Trm734 to modify position Cm32 and Gm34, respectively. Mutant strains lacking trm7 are sick and more sensitive to paromomycin, indicating Trm7 is crucial for cell translation. Still, the role of Cm32 and Gm34 modifications in translation remains unclear. Due to wobble pairing, tRNAPhe decodes both UUU and UUC codons. The RNA-ID reporter system can help study the dependence of UUU and UUC codons on Cm32 and Gm34, by allowing bidirectional expression of red fluorescence proteins (RFP) and green fluorescence proteins (GFP). Codons to be tested are inserted in front of GFP, allowing translation analysis in cells by their GFP/RFP ratio. We are currently working to integrate the reporter into trm734Δ and trm732Δ mutants to perform the experiments.

Investigation of 2’-O-methylation function on tRNAPhe

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