title: An alternative model for the role of RP2 protein in flagellum assembly in the African trypanosome.
abstract: The tubulin cofactor C domain-containing protein TbRP2 is a basal body (centriolar) protein essential for axoneme formation in the flagellate protist Trypanosoma brucei, the causal agent of African sleeping sickness. Here, we show how TbRP2 is targeted and tethered at mature basal bodies and provide novel insight into TbRP2 function. Regarding targeting, understanding how several hundred proteins combine to build a microtubule axoneme is a fundamental challenge in eukaryotic cell biology. We show that basal body localization of TbRP2 is mediated by twinned, N-terminal TOF (TON1, OFD1, and FOP) and LisH motifs, motifs that otherwise facilitate localization of only a few conserved proteins at microtubule-organizing centers in animals, plants, and flagellate protists. Regarding TbRP2 function, there is a debate as to whether the flagellar assembly function of specialized, centriolar tubulin cofactor C domain-containing proteins is processing tubulin, the major component of axonemes, or general vesicular trafficking in a flagellum assembly context. Here we report that TbRP2 is required for the recruitment of T. brucei orthologs of MKS1 and MKS6, proteins that, in animal cells, are part of a complex that assembles at the base of the flagellum to regulate protein composition and cilium function. We also identify that TbRP2 is detected by YL1/2, an antibody classically used to detect α-tubulin. Together, these data suggest a general processing role for TbRP2 in trypanosome flagellum assembly and challenge the notion that TbRP2 functions solely in assessing tubulin "quality" prior to tubulin incorporation into the elongating axoneme.
authors.affiliation: From the Faculty of Health and Medicine, Biomedical and Life Sciences, Lancaster University, Lancaster LA1 4YQ, United Kingdom.
authors.forename: Jane
authors.initials: J
authors.lastname: Andre
authors.forename: Louise
authors.initials: L
authors.lastname: Kerry
authors.forename: Xin
authors.initials: X
authors.lastname: Qi
authors.forename: Erica
authors.initials: E
authors.lastname: Hawkins
authors.forename: Kristina
authors.initials: K
authors.lastname: Drizyte
authors.forename: Michael L
authors.initials: ML
authors.lastname: Ginger
authors.forename: Paul G
authors.initials: PG
authors.lastname: McKean
mesh_terms: D020816:Amino Acid Motifs; D054468:Axoneme / Q000235:genetics / Q000378:metabolism*; D005407:Flagella / Q000235:genetics / Q000378:metabolism*; D005408:Flagellin / Q000235:genetics / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000378:metabolism*; D014404:Tubulin / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D014404:Tubulin; D005408:Flagellin
keywords: Axoneme; Centriole; Cilia; GTPase-activating Protein; Protein Targeting; Trypanosoma Brucei; Tubulin