Plasmodium falciparum PfMyoF contains a Rab-like tail domain and associates with perinuclear membrane trafficking proteins.

Members of the myosin superfamily are found in all eukaryotes, including Plasmodium falciparum, the parasite that causes the majority of severe malaria. The P. falciparum genome encodes six myosins, but apart from PfMyoA and B, these remain largely uncharacterised. Here, we characterise PfMyoF, a class XXII myosin, using structural prediction, biochemical assays, and imaging. We show that PfMyoF is a plus-end-directed, processive motor with a long neck region with capacity to bind up to six copies of the calmodulin homologue PfCaM. The PfMyoF tail contains predicted WD40 and Rab-like domains that have not been identified in any other eukaryotic myosin class. Pull-down experiments identify PfMyoF interactions with trafficking proteins, including the vesicle marker PfRab18. Expansion and immunoelectron microscopy reveal that PfMyoF localises to a perinuclear membrane compartment and transient knockdown using the glmS ribozyme system impairs growth, potentially indicating an important function during asexual replication. Our findings define PfMyoF as the first myosin with a Rab-like domain and highlight its potential role in membrane trafficking pathways during the pathogenic blood stages of parasite development.