N6-methyladenosine modification of messenger RNA (mRNA) was recently identified as an important genetic regulatory mechanism across eukarya. In the fungi, S. cerevisiae, Ime4p serves as the 6-methyladenosine methyltransferase that acts on mRNAs. In higher organisms a heterodimer of the METTL3 and METTL14 proteins is necessary for mRNA methylation and splice variants exist for METTL3. Because Ime4p directed methylation of mRNA is a simpler system and high-quality, genome-wide maps of N6-methyladenosine sites in S. cerevisiae exist, we are exploring Ime4p as a model system for N6-methyladenosine mRNA modification. We report our progress on cloning of IME4 gene constructs for recombinant expression and purification in E. coli to develop in vitro 6-methyladenosine modification assays.

Cloning of IME4 to establish a new model system to probe N6-methyladenosine modification of mRNA

253rd National Meeting (2017)

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