Poster Presentation Lorne Infection and Immunity 2019

Investigating the epitranscriptome of Plasmodium parasites (#120)

Amy J Distiller 1 , V Vern Lee 1 , Emily M Crisafulli 1 , Michael Duffy 2 , Stuart A Ralph 1
  1. Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Vic, Australia
  2. School of Biosciences, The University of Melbourne, Parkville, Vic, Australia

Recent years have seen an increasing identification and awareness of post-transcriptional modifications of RNA. Modifications have long been known to be important in diverse non-coding RNAs, but post-transcriptional modifications of mRNA are now also recognised as playing an important role in regulation of gene expression. One of the most abundant mRNA modifications is N6-methyladenosine (m6A), the presence of which guides mRNA metabolism including maturation, nuclear export, translation and decay. In model eukaryotes, m6A is produced by the action of METTL-family methyltransferases, and these enzymes play a key role in cellular differentiation and development. We identified several METTL methyltransferases in the Plasmodium genome, and are now investigating RNA modifications in the transcriptome of Plasmodium parasites. We have studied and characterised two methyltransferases, Mettl3-like proteins in P. falciparum through localisation, overexpression experiments and knock-sideways studies. We will investigate the influence of Mettl3-like proteins on splicing patterns in the parasite, and test the role of these modifications on proliferation and differentiation. We have employed nanopore direct-RNA sequencing to analyse transcriptome patterns in Plasmodium, and detected modified bases from MinION nanopore sequence traces. We will use this sequence to characterise m6A modifications in wild type parasites as well as parasites with disrupted METTL methyltransferases.