title: Biogenesis of the crystalloid organelle in Plasmodium involves microtubule-dependent vesicle transport and assembly.
abstract: Malaria parasites possess unique subcellular structures and organelles. One of these is the crystalloid, a multivesicular organelle that forms during the parasite's development in vector mosquitoes. The formation and function of these organelles remain poorly understood. A family of six conserved and modular proteins named LCCL-lectin adhesive-like proteins (LAPs), which have essential roles in sporozoite transmission, localise to the crystalloids. In this study we analyse crystalloid formation using transgenic Plasmodium berghei parasites expressing GFP-tagged LAP3. We show that deletion of the LCCL domain from LAP3 causes retarded crystalloid development, while knockout of LAP3 prevents formation of the organelle. Our data reveal that the process of crystalloid formation involves active relocation of endoplasmic reticulum-derived vesicles to common assembly points via microtubule-dependent transport. Inhibition of microtubule-dependent cargo transport disrupts this process and replicates the LCCL domain deletion mutant phenotype in wildtype parasites. These findings provide the first clear insight into crystalloid biogenesis, demonstrating a fundamental role for the LAP family in this process, and identifying the crystalloid and its formation as potential targets for malaria transmission control.
authors.affiliation: Pathogen Molecular Biology Department, Faculty of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom.
authors.forename: Sadia
authors.initials: S
authors.lastname: Saeed
authors.affiliation: Pathogen Molecular Biology Department, Faculty of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom.
authors.forename: Annie Z
authors.initials: AZ
authors.lastname: Tremp
authors.affiliation: Pathogen Molecular Biology Department, Faculty of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom. Electronic address: Johannes.Dessens@lshtm.ac.uk.
authors.forename: Johannes T
authors.initials: JT
authors.lastname: Dessens
mesh_terms: D000818:Animals; D005260:Female; D006801:Humans; D008288:Malaria / Q000469:parasitology*; D051379:Mice; D008870:Microtubules / Q000235:genetics / Q000378:metabolism*; D015388:Organelles / Q000235:genetics / Q000378:metabolism*; D010962:Plasmodium berghei / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D022161:Transport Vesicles / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins
keywords: Cargo transport; Crystalloid; LCCL protein; Plasmodium berghei; Transgenic parasite