The rhoptry of the malaria parasite Plasmodium falciparum is an unusual secretory organelle that is thought to be related to secretory lysosomes in higher eukaryotes. Rhoptries contain an extensive collection of proteins that participate in host cell invasion and in the formation of the parasitophorous vacuole, but little is known about sorting signals required for rhoptry protein targeting. Using green fluorescent protein chimeras and in vitro pull-down assays, we performed an analysis of the signals required for trafficking of the rhoptry protein RAP1. We provide evidence that RAP1 is escorted to the rhoptry via an interaction with the glycosylphosphatidyl inositol-anchored rhoptry protein RAMA. Once within the rhoptry, RAP1 contains distinct signals for localisation within a sub-compartment of the organelle and subsequent transfer to the parasitophorous vacuole after invasion. This is the first detailed description of rhoptry trafficking signals in Plasmodium.
The malaria parasite Plasmodium falciparum is a eukaryotic organism with multiple membrane bound organelles with discrete functions. The rhoptry is an unusual secretory organelle that participates in host cell invasion and the formation of a specialised vacuole that the parasite occupies during the intracellular part of its lifecycle. Rhoptries contain an extensive collection of proteins, but little is known about how these proteins are trafficked to their destination. Understanding determinants of rhoptry protein trafficking will help us to identify novel rhoptry proteins, and may provide targets for therapeutic intervention. In the current study, we focussed on the trafficking of the rhoptry protein RAP1. By making parasites that express regions of RAP1 fused to Green Fluorescent Protein (GFP), we were able to map in detail the domains of RAP1 that are necessary for correct trafficking. We also provide evidence that RAP1 is targeted to rhoptries via its interaction with another rhoptry protein, RAMA. This is the first detailed description of rhoptry trafficking signals in Plasmodium.