title: Proteomic identification of novel cytoskeletal proteins associated with TbPLK, an essential regulator of cell morphogenesis in Trypanosoma brucei.
abstract: Trypanosoma brucei is the causative agent of African sleeping sickness, a devastating disease endemic to sub-Saharan Africa with few effective treatment options. The parasite is highly polarized, including a single flagellum that is nucleated at the posterior of the cell and adhered along the cell surface. These features are essential and must be transmitted to the daughter cells during division. Recently we identified the T. brucei homologue of polo-like kinase (TbPLK) as an essential morphogenic regulator. In the present work, we conduct proteomic screens to identify potential TbPLK binding partners and substrates to better understand the molecular mechanisms of kinase function. These screens identify a cohort of proteins, most of which are completely uncharacterized, which localize to key cytoskeletal organelles involved in establishing cell morphology, including the flagella connector, flagellum attachment zone, and bilobe structure. Depletion of these proteins causes substantial changes in cell division, including mispositioning of the kinetoplast, loss of flagellar connection, and prevention of cytokinesis. The proteins identified in these screens provide the foundation for establishing the molecular networks through which TbPLK directs cell morphogenesis in T. brucei.
authors.affiliation: Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912.
authors.forename: Michael R
authors.initials: MR
authors.lastname: McAllaster
authors.affiliation: Max F. Perutz Laboratories, Department of Medical Biochemistry, Medical University of Vienna, 1030 Vienna, Austria.
authors.forename: Kyojiro N
authors.initials: KN
authors.lastname: Ikeda
authors.affiliation: Max F. Perutz Laboratories, Center for Molecular Biology, University of Vienna, 1030 Vienna, Austria.
authors.forename: Ana
authors.initials: A
authors.lastname: Lozano-Núñez
authors.affiliation: Max F. Perutz Laboratories, Mass Spectrometry Facility, University of Vienna, 1030 Vienna, Austria.
authors.forename: Dorothea
authors.initials: D
authors.lastname: Anrather
authors.affiliation: Max F. Perutz Laboratories, Mass Spectrometry Facility, University of Vienna, 1030 Vienna, Austria.
authors.forename: Verena
authors.initials: V
authors.lastname: Unterwurzacher
authors.affiliation: Max F. Perutz Laboratories, Mass Spectrometry Facility, University of Vienna, 1030 Vienna, Austria.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gossenreiter
authors.affiliation: Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912.
authors.forename: Jenna A
authors.initials: JA
authors.lastname: Perry
authors.affiliation: Department of Biological and Medical Sciences, Faculty of Health and Life Science, Oxford Brookes University, Oxford OX3 0BP, United Kingdom.
authors.forename: Robbie
authors.initials: R
authors.lastname: Crickley
authors.affiliation: Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912.
authors.forename: Courtney J
authors.initials: CJ
authors.lastname: Mercadante
authors.affiliation: Department of Biological and Medical Sciences, Faculty of Health and Life Science, Oxford Brookes University, Oxford OX3 0BP, United Kingdom.
authors.forename: Sue
authors.initials: S
authors.lastname: Vaughan
authors.affiliation: Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912 Christopher_deGraffenried@Brown.edu.
authors.forename: Christopher L
authors.initials: CL
authors.lastname: de Graffenried
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D048749:Cytokinesis; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism; D009024:Morphogenesis; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D000097869:Polo-Like Kinase 1
chemical_list: D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D015800:Protozoan Proteins; D000097869:Polo-Like Kinase 1