A Tale of Two Parasites: A Glimpse into the RNA Methylome of Patient-derived P. falciparum and P. vivax isolates

Understanding the molecular mechanisms of the malarial parasites in hosts is crucial for developing effective treatments. Epitranscriptomic research on pathogens has unveiled the significance of RNA methylation in gene regulation and pathogenesis. We present the first report investigating methylation patterns determined using Nanopore Direct RNA Sequencing, to single-base resolution across RNA of P. falciparum and P. vivax from Malaria patients with hepatic dysfunction. Distinct RNA methylation profiles of N6-methyladenosine and 5-methylcytosine in the parasites suggest species-specific regulatory mechanisms. These modifications are unevenly present in the annotated regions of the mRNA, potentially influencing mRNA export and translation. Methylated antisense transcripts, splicing events and modified transcripts originating from apicoplast and mitochondrial genomes have been detected. We observe a striking overlap of differential methylation of isoforms with either of the modifications in both species. The correlation of these regulatory layers will decipher the post-transcriptional environment of malaria parasites in vivo and elucidate their inherent proteome plasticity.