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Flagellar motility is required for the viability of the bloodstream trypanosome

Nature volume 440, pages 224–227 (2006)Cite this article

Abstract

The 9 + 2 microtubule axoneme of flagella and cilia represents one of the most iconic structures built by eukaryotic cells and organisms. Both unity and diversity are present among cilia and flagella on the evolutionary as well as the developmental scale. Some cilia are motile, whereas others function as sensory organelles and can variously possess 9 + 2 and 9 + 0 axonemes and other associated structures1. How such unity and diversity are reflected in molecular repertoires is unclear. The flagellated protozoan parasite Trypanosoma brucei is endemic in sub-Saharan Africa, causing devastating disease in humans and other animals2. There is little hope of a vaccine for African sleeping sickness and a desperate need for modern drug therapies3. Here we present a detailed proteomic analysis of the trypanosome flagellum. RNA interference (RNAi)-based interrogation of this proteome provides functional insights into human ciliary diseases and establishes that flagellar function is essential to the bloodstream-form trypanosome. We show that RNAi-mediated ablation of various proteins identified in the trypanosome flagellar proteome leads to a rapid and marked failure of cytokinesis in bloodstream-form (but not procyclic insect-form) trypanosomes, suggesting that impairment of flagellar function may provide a method of disease control. A postgenomic meta-analysis, comparing the evolutionarily ancient trypanosome with other eukaryotes including humans, identifies numerous trypanosome-specific flagellar proteins, suggesting new avenues for selective intervention.

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Figure 1: The T. brucei flagellar proteome.
Figure 2: Biological verification of the T. brucei flagellar proteome.
Figure 3: Flagellar motility is essential for bloodstream trypanosomes.

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Acknowledgements

We thank the genome centres, particularly GeneDB at the Sanger Institute, and our laboratory colleagues, especially S. Gordon and P. Taylor, for access to unpublished data and constructive discussions. This work was funded by the Biotechnology and Biological Sciences Research Council, the Wellcome Trust, the Royal Society and the Edward P. Abraham Trust. M.L.G. is a Royal Society University Research Fellow; K.G. is a Wellcome Trust Principal Research Fellow. Author Contributions The first six authors contributed equally to this work and are listed alphabetically. Individual contributions were as follows: R.B. proteomics and bioinformatic analysis; H.R.D. proteomics, bioinformatic and functional analyses; H.F. and S.G. functional analysis; S.R.H. proteomics; N.P. proteomics, bioinformatic and functional analyses.

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Author notes

  1. Richard Broadhead, Helen R. Dawe, Helen Farr, Samantha Griffiths, Sarah R. Hart and Neil Portman: *These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biological Sciences, Lancaster University, LA1 4YQ, Lancaster, UK

    Richard Broadhead & Paul G. McKean

  2. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE, Oxford, UK

    Helen R. Dawe, Helen Farr, Samantha Griffiths, Neil Portman, Michael K. Shaw, Michael L. Ginger & Keith Gull

  3. Michael Barber Centre for Mass Spectrometry, School of Chemistry, University of Manchester, Sackville Street, M60 1QD, Manchester, UK

    Sarah R. Hart & Simon J. Gaskell

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Correspondence to Keith Gull.

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Supplementary information

Supplementary Notes 1

This file contains and introduction and preface to the Supplementary Information and the Supplementary Figure Legends. (DOC 23 kb)

Supplementary Methods

This file contains additional details of the methods used in this study. (DOC 47 kb)

Supplementary Notes 2

List of the key proteins in this study. (DOC 23 kb)

Supplementary Figure 1

The T. brucei Flagellar Proteome (TbFP): Table showing the proteins making up the TbFP with the number of unique peptides identifying each protein and the percentage sequence coverage this represents. Predicted protein characteristics and comparative bioinformatics conducted against an informative set of model organisms are also shown. (PDF 150 kb)

Supplementary Figure 2

Ciliary disease genes and in the TbFP a. Table showing TbFP proteins with an associated human homologue that has been shown to be the causative agent of a disease with known ciliary links. b. Candidate human disease genes in the TbFP (PDF 24 kb)

Supplementary Figure 3

Flagellar motility is essential for bloodstream trypanosomes. Table summarizing the phenotypes observed from biological validation of the TbFP by RNAi of bloodstream form trypanosomes. Transmission EMs showing phenotypes of RNAi-induced PFR2, MBO2, DIGIT and TAX1 in bloodstream form trypanosomes. (PDF 93 kb)

Supplementary Figure 4

Analysis by tomato lectin and VSG antibody shows that the pocket is functional to varying degrees in PFR2 RNAi-induced bloodstream form cells. Immunofluorescence images showing flagellar pocket functionality in bloodstream form PFR2 RNAi-induced cells. (PDF 2621 kb)

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Broadhead, R., Dawe, H., Farr, H. et al. Flagellar motility is required for the viability of the bloodstream trypanosome. Nature 440, 224–227 (2006). https://doi.org/10.1038/nature04541

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