Kinesin motor proteins in malaria. (360G-Wellcome-085945_Z_08_Z)

Kinesins are microtubule-based, ATP-dependent molecular motors that have many essential roles in eukaryotes including in cell division, cell motility and intracellular transport. Elimination of kinesin function often proves fatal because essential cell processes are compromised. The malaria genome encodes ten putative kinesin motors, about which little is known, and the proposed research will aim to characterise them biochemically, structurally and within the parasite. Key goals: Assessment of kinesin expression in parasites by RT-PCR Clone and express Plasmodium kinesins, particularly their motor domains which contain microtubule and ATP-binding sites. Begin with four kinesins with homologues in humans. Biochemical characterisation of microtubule-stimulated ATPase activity, providing insight into likely functions Visualisation of the kinesin motor-microtubule interaction by cryo-electron microscopy and 3D reconstruction Generate pseudo-atomic models of the motor-microt ubule interaction using homology models and dock these into the EM-derived molecular envelope Expression of GFP-tagged motors in vivo to determine their location and assess their possible roles in vivo Use of electron tomography to visualise the morphology of parasite microtubules and correlate this with kinesin localisation Combine in vitro and in vivo assays to set up drug screens to look for kinesin-specific inhibitors