The Plasmodium falciparum protein VCAP1 controls Maurer’s cleft morphology, knob architecture and PfEMP1 trafficking

McHugh, E.; Carmo, O.; Blanch, A.; Looker, O.; Liu, B.; Tiash, S.; Batinovic, S.; Low J.Y, A.; McMillan, P.; Tilley, L.; Dixon, M.

doi:10.1101/741033

bioRxiv · 10.1101/741033

The Plasmodium falciparum protein VCAP1 controls Maurer’s cleft morphology, knob architecture and PfEMP1 trafficking

McHugh, E.·Carmo, O.·Blanch, A.·Looker, O.·Liu, B.·Tiash, S.·Batinovic, S.·Low J.Y, A.·McMillan, P.·Tilley, L.·Dixon, M.

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Abstract

The malaria parasite, Plasmodium falciparum, traffics the virulence protein, P. falciparum erythrocyte membrane protein 1 (PfEMP1) to the surface of infected red blood cells (RBCs) via membranous organelles, known as the Maurers clefts. We developed a method for efficient enrichment of Maurers clefts and profiled the protein composition of this trafficking organelle. We identified 13 previously uncharacterised or poorly characterised Maurers cleft proteins. We generated transfectants expressing GFP-fusions of 7 proteins and confirmed their Maurers cleft location. Using co-immunoprecipitation and mass spectrometry we have generated a protein interaction map of proteins at the Maurers clefts. We identified two key clusters that may function in the loading and unloading of PfEMP1 into and out of the Maurers clefts. We focus on a putative PfEMP1 loading complex that includes the newly characterised virulence complex assembly protein 1 (VCAP1). Disruption of VCAP1 causes Maurers cleft fragmentation, aberrant knobs, ablation of PfEMP1 surface expression and loss of the PfEMP1 directed adhesion. {Delta}VCAP1 parasite lines have a growth advantage compared to wildtype parasites; and the infected RBCs are more deformable and more osmotically fragile.\n\nImportanceThe trafficking of the virulence antigen PfEMP1 and its presentation at the knob structures at the surface of parasite infected RBCs is central to severe adhesion related pathologies such as cerebral and placental malaria. This work adds to our understanding of how PfEMP1 is trafficked to the RBC membrane by defining the protein-protein interaction networks that function at the Maurers clefts controlling PfEMP1 loading and unloading. This work adds significantly to our understanding of virulence protein trafficking and will provide crucial knowledge that will be required to determine the mechanisms underpinning parasite driven host cell remodelling, parasite survival within the host and virulence mechanisms.

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McHugh, E., Carmo, O., Blanch, A., Looker, O., Liu, B., Tiash, S., Batinovic, S., Low J.Y, A., McMillan, P., Tilley, L., Dixon, M.. 2019-08-20. The Plasmodium falciparum protein VCAP1 controls Maurer’s cleft morphology, knob architecture and PfEMP1 trafficking. https://doi.org/10.1101/741033

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