Oral Presentation Australasian RNA Biology and Biotechnology Association 2024 Conference

mRNA base modifications in the malaria parasite Plasmodium falciparum impact transcript stability and translational efficiency (109112)

Stuart A Ralph 1 , Lakvin Fernando 1 , Joshua Levendis 1 , Amy Distiller 1 , Emma McHugh 1
  1. Biochemistry and Pharmacology, The University Of Melbourne, Parkville, VIC, Australia

RNA can be chemically modified at a gene-specific level, and RNA modifications have been central to the success of COVID-19 mRNA vaccines. Despite the importance of these modifications the role of the most abundant RNA modifications remains unclear. We used Nanopore sequencing to interrogate the epitranscriptome of Plasmodium falciparum. Plasmodium has a small transcriptome with only 3000 genes expressed in any one life stage, making it ideally suited for whole transcriptome analyses. We mapped transcriptomes with individual Nanopore flow cells, and interrogated the prevalence and position of RNA modifications. Plasmodium transcripts are the most adenosine rich in any known eukaryote, and N6-Methyladenosine (m6A) is particularly abundant in Plasmodium mRNA. We perturbed the writing and reading of this epitranscriptome code by inducibly knocking down the methyltransferase that lays down m6A in mRNA and the reader proteins that detect and decode m6A containing transcripts. These perturbations impact mRNA and protein abundance, pointing to nuanced roles for m6A in RNA stability and translational efficiency. The contextual importance of m6A sites remains enigmatic, and we are investigating the impact of methylation position and density within transcripts to better understand how m6A can be effectively exploited as a biotechnology tool.