Molecular architecture of glideosome and nuclear F-actin inPlasmodium falciparum

Abstract

Actin-based motility is required for the transmission of malaria sporozoites. While this has been shown biochemically, filamentous actin has remained elusive and has to date never been directly visualised inside the parasite. Using focused ion beam milling and electron cryo-tomography, we studied dynamic actin filaments in unperturbedPlasmodium falciparumcells for the first time. This allowed us to dissect the assembly, path and fate of actin filaments during parasite gliding and determine a complete 3D model of F-actin within sporozoites. We show that within the cell, actin assembles into micrometre long filaments, much longer than observed inin vitrostudies. After their assembly at the parasite’s apical end, actin filaments continue to grow as they are transported down the cell as part of the glideosome machinery, and are disassembled at the basal end in a rate-limiting step. Large pores in the IMC, constrained to the basal end, may facilitate actin exchange between the pellicular space and the cytosol for its recycling and maintenance of directional actin flow for efficient gliding. The data also reveal striking and extensive actin bundles in the nucleus. Implications of these structures for motility and transmission are discussed.