Gujarati, S.; Gupta, D.; Kochar, S. K.; Kochar, D. K.; Das, A.

The malaria parasite mitochondrial proteins are critical targets for antimalarial drugs, however, the emergence of drug resistance against the existing protein targets necessitates novel therapeutic approaches. In this study, using custom-designed, multi-probe microarrays and pooled RNA sequencing experiments in patient samples of Plasmodium falciparum, we identified disease-specific sense and Natural Antisense Transcripts (NATs) encoded by the mitochondrial genome, along with those encoded in the nucleus and targeted to the parasite mitochondria. These findings span two clinical manifestations of severe malaria - hepatic dysfunction (HD) and cerebral malaria (CM). Although mitochondrial activity is known to be reduced in blood-stage malaria, upregulation of genes linked to the TCA cycle in the CM cluster indicates disrupted mitochondrial bioenergetics which might contribute to severe disease pathogenesis. Profiling of sense and antisense mitochondrial transcripts reveal a striking correlated expression of sense-antisense transcript pairs in both the disease manifestations, indicating a potential regulatory role of NATs in mitochondrial function. This study provides direct evidence for the origin of NATs from the mitochondrial genome. It further provides a dynamic gene regulation profile of key sense and antisense transcripts responsible for mitochondrial translation and bioenergetics. We propose these differentially regulated NATs against protein-coding genes specific to the observed disease severity as potential novel targets to disrupt key mitochondrial functions of the parasite.