Molecular mechanisms of sporogonic development in malaria parasites

Objectives

This project aims to study the mechanisms of sporogonic development focussing on the role of palmitoylation in crystalloid biogenesis and function. Using the rodent malaria parasite Plasmodium berghei, this study will dissect the substrate range, spatiotemporal dynamics and mechanisms of DHHC10-mediated palmitoylation in crystalloids. The objectives are: (1) Using acyl-biotin-exchange (ABE) and label-free quantitative mass spectrometry (LFQMS), the ookinete palmitome will be determined of DHHC10-positive and DHHC10-negative parasite, allowing the identification of DHHC10 substrates and the crystalloid palmitome; (2) Proteins that interact with DHHC10 (either subunits of a DHHC10 protein complex, or its substrates) will be isolated by GFP pull-down using ookinetes expressing GFP-tagged DHHC10, and identified by LFQMS. In a second approach, interactors for DHHC10 will be identified by BioID using ookinetes that express DHHC10 fused to a biotin ligase; (3) New candidate crystalloid proteins identified will be validated using fluorescent protein tagging and gene knockout in transgenic parasite lines.