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African trypanosomes and related kinetoplastid parasites selectively traffic specific membrane proteins to the flagellar membrane, but the mechanisms for this trafficking are poorly understood. We show here that KHARON, a protein originally identified in Leishmania parasites, interacts with a putative trypanosome calcium channel and is required for its targeting to the flagellar membrane. KHARON is located at the base of the flagellar axoneme, where it likely mediates targeting of flagellar membrane proteins, but is also on the subpellicular microtubules and the mitotic spindle. Hence, KHARON is probably a multifunctional protein that associates with several components of the trypanosome cytoskeleton. RNA interference-mediated knockdown of KHARON mRNA results in failure of the calcium channel to enter the flagellar membrane, detachment of the flagellum from the cell body, and disruption of mitotic spindles. Furthermore, knockdown of KHARON mRNA induces a lethal failure of cytokinesis in both bloodstream (mammalian host) and procyclic (insect vector) life cycle stages, and KHARON is thus critical for parasite viability.

Original publication

DOI

10.1074/jbc.M116.739235

Type

Journal article

Journal

J Biol Chem

Publication Date

16/09/2016

Volume

291

Pages

19760 - 19773

Keywords

Trypanosoma brucei, calcium channel, cytoskeleton, membrane protein, mitotic spindle, trafficking, Calcium Channels, Cell Membrane, Cytokinesis, Cytoskeletal Proteins, Flagella, Gene Knockdown Techniques, Leishmania, Protozoan Proteins, Spindle Apparatus, Trypanosoma brucei brucei