title: An analogue-sensitive approach identifies basal body rotation and flagellum attachment zone elongation as key functions of PLK in Trypanosoma brucei.
abstract: Polo-like kinases are important regulators of cell division, playing diverse roles in mitosis and cytoskeletal inheritance. In the parasite Trypanosoma brucei, the single PLK homologue TbPLK is necessary for the assembly of a series of essential organelles that position and adhere the flagellum to the cell surface. Previous work relied on RNA interference or inhibitors of undefined specificity to inhibit TbPLK, both of which have significant experimental limitations. Here we use an analogue-sensitive approach to selectively and acutely inhibit TbPLK. T. brucei cells expressing only analogue-sensitive TbPLK (TbPLK(as)) grow normally, but upon treatment with inhibitor develop defects in flagellar attachment and cytokinesis. TbPLK cannot migrate effectively when inhibited and remains trapped in the posterior of the cell throughout the cell cycle. Using synchronized cells, we show that active TbPLK is a direct requirement for the assembly and extension of the flagellum attachment zone, which adheres the flagellum to the cell surface, and for the rotation of the duplicated basal bodies, which positions the new flagellum so that it can extend without impinging on the old flagellum. This approach should be applicable to the many kinases found in the T. brucei genome that lack an ascribed function.
authors: Max F. Perutz Laboratories, Center for Molecular Biology, University of Vienna, 1030 Vienna, Austria.
authors: Ana
authors: A
authors: Lozano-Núñez
authors: Kyojiro N
authors: KN
authors: Ikeda
authors: Thomas
authors: T
authors: Sauer
authors: Christopher L
authors: CL
authors: de Graffenried
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D016054:DNA, Protozoan / Q000235:genetics / Q000378:metabolism; D005407:Flagella / Q000187:drug effects / Q000201:enzymology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011687:Purines / Q000494:pharmacology*; D061987:Sf9 Cells; D018411:Spodoptera; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014346:Trypanosoma brucei brucei / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics
chemical_list: C584364:3MB-PP1; D016054:DNA, Protozoan; D015800:Protozoan Proteins; D011687:Purines; D014344:Trypanocidal Agents; D017346:Protein Serine-Threonine Kinases