An in silico down-scaling approach uncovers novel constituents of the Plasmodium-containing vacuole.

During blood stage development the malaria parasite resides in a membrane-bound compartment, termed the parasitophorous vacuole (PV). The reasons for this intravacuolar life style and the molecular functions of this parasite-specific compartment remain poorly defined, which is mainly due to our limited knowledge about the molecular make-up of this unique niche. We used an in silico down-scaling approach to select for Plasmodium-specific candidates that harbour signatures of PV residency. Live co-localisation of five endogenously tagged proteins confirmed expression in the PV of Plasmodium berghei blood and liver stages. ER retention was ruled out by addition of the respective carboxyterminal tetrapeptides to a secreted reporter protein. Although all five PV proteins are highly expressed, four proved to be dispensable for parasite development in the mammalian and mosquito host, as revealed by targeted gene deletion. In good agreement with their redundant roles, the knockout parasites displayed no detectable deficiencies in protein export, sequestration, or PV morphology. Together, our approach improved the catalogue of the Plasmodium PV proteome and provides experimental genetics evidence for functional redundancy of several PV proteins.