The P. falciparum orthologue of Male Development Protein 3 is a Male-associated regulator of translation initiation or enhancement [CLIP-Seq]

Sex determination and development of Plasmodium gametocytes is critical to the transmission of malaria parasites and involves a complex interplay of multiple molecular factors. Here, we characterized the P. falciparum gene syntenic to P. berghei MD3- pf3d7_0315600. This protein contains a non-classical CCCH zinc-finger domain and Lysine-Proline rich domain associated with ribosome receptors, suggesting a role in mRNA regulation. We investigated the function of PfMD3-3xHA using in vivo using crosslinked RNA-Protein interaction capture followed by RNA-sequencing (CLIP-seq) to determine its RNA targets in both early (stage II) and late (stage IV-V) development of gametocytes. We found that these bound mRNA targets show decreased protein levels in a genetic knockout of PfMD3, suggesting a role for PfMD3 in translational processing. We then propose that PfMD3 functions as a Male Associated Regulator of Translation Initiation or Enhancement (MARTIE) in the human malaria parasite P. falciparum. Overall design: Crosslinked RNA-Protein interaction capture followed by RNA-sequencing (CLIP-seq) for PfMD3-3xHA in Plasmodium falciparum NF54e parasites in stage II-III and stage IV-V gametocytes (4 biological replicates each)