title: Flagellar motility contributes to cytokinesis in Trypanosoma brucei and is modulated by an evolutionarily conserved dynein regulatory system.
abstract: The flagellum of Trypanosoma brucei is a multifunctional organelle with critical roles in motility and other aspects of the trypanosome life cycle. Trypanin is a flagellar protein required for directional cell motility, but its molecular function is unknown. Recently, a trypanin homologue in Chlamydomonas reinhardtii was reported to be part of a dynein regulatory complex (DRC) that transmits regulatory signals from central pair microtubules and radial spokes to axonemal dynein. DRC genes were identified as extragenic suppressors of central pair and/or radial spoke mutations. We used RNA interference to ablate expression of radial spoke (RSP3) and central pair (PF16) components individually or in combination with trypanin. Both rsp3 and pf16 single knockdown mutants are immotile, with severely defective flagellar beat. In the case of rsp3, this loss of motility is correlated with the loss of radial spokes, while in the case of pf16 the loss of motility correlates with an aberrant orientation of the central pair microtubules within the axoneme. Genetic interaction between trypanin and PF16 is demonstrated by the finding that loss of trypanin suppresses the pf16 beat defect, indicating that the DRC represents an evolutionarily conserved strategy for dynein regulation. Surprisingly, we discovered that four independent mutants with an impaired flagellar beat all fail in the final stage of cytokinesis, indicating that flagellar motility is necessary for normal cell division in T. brucei. These findings present the first evidence that flagellar beating is important for cell division and open the opportunity to exploit enzymatic activities that drive flagellar beat as drug targets for the treatment of African sleeping sickness.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, 609 Charles E. Young Dr., Los Angeles, CA 90095, USA.
authors.forename: Katherine S
authors.initials: KS
authors.lastname: Ralston
authors.forename: Alana G
authors.initials: AG
authors.lastname: Lerner
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Diener
authors.forename: Kent L
authors.initials: KL
authors.lastname: Hill
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D017124:Conserved Sequence; D048749:Cytokinesis / Q000502:physiology*; D004398:Dyneins / Q000235:genetics / Q000502:physiology*; D019143:Evolution, Molecular; D005407:Flagella / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation; D015800:Protozoan Proteins / Q000235:genetics / Q000502:physiology*; D016415:Sequence Alignment; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000502:physiology*
chemical_list: D015800:Protozoan Proteins; C107172:T lymphocyte triggering factor, Trypanosoma brucei; D004398:Dyneins