Abstract
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.
Highlights
Malaria is a disease that causes severe morbidity, mortality and socio-economic hardship in tropical and sub-tropical areas of Africa, South America and Asia
To more precisely define these targeting signals, we used constructs consisting of regions of rhoptry associated protein 1 (RAP1) fused to green fluorescent protein (GFP)
The expression of RAP1-Green Fluorescent Protein (GFP) chimeras was driven by an inducible promoter with a pattern of expression similar to merozoite surface protein 2 (MSP2) [26]
Summary
Malaria is a disease that causes severe morbidity, mortality and socio-economic hardship in tropical and sub-tropical areas of Africa, South America and Asia. Plasmodium falciparum causes the most serious form of the disease and is responsible for more than 2 million deaths annually [1,2,3]. The invasion is mediated by a set of molecules distributed on the parasite surface and within specialised apical secretory organelles. Regulated secretion from these organelles allows the parasite to adhere to an appropriate target cell, invade and induce the formation of a specialised parasitophorous vacuole (PV) in which it subsequently resides (reviewed in [5])
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