Plasmodium spp. are the single-celled, eukaryotic causative agents of malaria, with P. falciparum causing the most severe form of the disease in humans. This parasite has a small, relatively well annotated genome of around 5,000 genes. The regulation of expression of these genes in the pathogenic stage – the intra-erythrocytic developmental cycle – underlies key mechanisms of pathogenesis.
Plasmodium falciparum is known to contain many mRNA modifications and N6-methyladenosine (m6A) is the most abundant among these. In other eukaryotes, m6A is deposited co-transcriptionally, therefore can influence mRNA throughout its lifetime.
The m6A writer’s core in P. falciparum is related to other known eukaryotic m6A writers, consisting of the WTAP adaptor and orthologs of METTL3 and METTL14. The METTL3 ortholog, is essential during the intra-erythrocytic development, and m6A is likely an important regulator of gene expression at this stage of the parasites’ lifecycle. We use knock-sideways to induce a functional knockdown of the METTL3 ortholog to study the effects of m6A in P. falciparum.
Our results suggests that m6A affects the cytoplasmic stability of mRNA, a potential mechanism of repressing their expression. We also investigate the role that m6A plays in mRNA export and translational efficiency.
The relatively small genome of P. falciparum makes transcriptomic and proteomic studies more feasible. As the m6A machinery in P. falciparum is related to that of humans, insights gained this work can be applied to work on human mRNA biology, RNA viruses that infect eukaryotes and mRNA vaccines.