The cell shape of Trypanosoma brucei is influenced by flagellum-to-cell-body attachment through a specialised structure - the flagellum attachment zone (FAZ). T. brucei exhibits numerous morphological forms during its life cycle and, at each stage, the FAZ length varies. We have analysed FLAM3, a large protein that localises to the FAZ region within the old and new flagellum. Ablation of FLAM3 expression causes a reduction in FAZ length; however, this has remarkably different consequences in the tsetse procyclic form versus the mammalian bloodstream form. In procyclic form cells FLAM3 RNAi results in the transition to an epimastigote-like shape, whereas in bloodstream form cells a severe cytokinesis defect associated with flagellum detachment is observed. Moreover, we demonstrate that the amount of FLAM3 and its localisation is dependent on ClpGM6 expression and vice versa. This evidence demonstrates that FAZ is a key regulator of trypanosome shape, with experimental perturbations being life cycle form dependent. An evolutionary cell biology explanation suggests that these differences are a reflection of the division process, the cytoskeleton and intrinsic structural plasticity of particular life cycle forms.

Faculty of Health and Medicine Division of Biomedical and Life Sciences Lancaster University Lancaster LA1 4YQ UK

Faculty of Sciences University of South Bohemia České Budějovice 37005 Czech Republic

Faculty of Sciences University of South Bohemia České Budějovice 37005 Czech Republic Canadian Institute for Advanced Research Toronto Ontario Canada M5G 1Z8

Sir William Dunn School of Pathology University of Oxford Oxford OX1 3RE UK

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Briggs L. J., McKean P. G., Baines A., Moreira-Leite F., Davidge J., Vaughan S. and Gull K. (2004). The flagella connector of PubMed DOI

Changmai P., Horáková E., Long S., Černotíková-Stříbrná E., McDonald L. M., Bontempi E. J. and Lukeš J. (2013). Both human ferredoxins equally efficiently rescue ferredoxin deficiency in PubMed DOI

Davidge J. A., Chambers E., Dickinson H. A., Towers K., Ginger M. L., McKean P. G. and Gull K. (2006). Trypanosome IFT mutants provide insight into the motor location for mobility of the flagella connector and flagellar membrane formation. PubMed DOI

Fields S. D., Conrad M. N. and Clarke M. (1998). The PubMed

Gao J., Schatton D., Martinelli P., Hansen H., Pla-Martin D., Barth E., Becker C., Altmueller J., Frommolt P., Sardiello M. et al. (2014). CLUH regulates mitochondrial biogenesis by binding mRNAs of nuclear-encoded mitochondrial proteins. PubMed DOI PMC

García-Salcedo J. A., Pérez-Morga D., Gijón P., Dilbeck V., Pays E. and Nolan D. P. (2004). A differential role for actin during the life cycle of PubMed DOI PMC

Gluenz E., Povelones M. L., Englund P. T. and Gull K. (2011). The kinetoplast duplication cycle in PubMed DOI PMC

Hayes P., Varga V., Olego-Fernandez S., Sunter J., Ginger M. L. and Gull K. (2014). Modulation of a cytoskeletal calpain-like protein induces major transitions in trypanosome morphology. PubMed DOI PMC

Hoare C. A. and Wallace F. G. (1966). Developmental stages of trypanosomatid flagellates: a new terminology. DOI

Kelly S., Reed J., Kramer S., Ellis L., Webb H., Sunter J., Salje J., Marinsek N., Gull K., Wickstead B. et al. (2007). Functional genomics in PubMed DOI PMC

Kohl L., Sherwin T. and Gull K. (1999). Assembly of the paraflagellar rod and the flagellum attachment zone complex during the PubMed DOI

Kohl L., Robinson D. and Bastin P. (2003). Novel roles for the flagellum in cell morphogenesis and cytokinesis of trypanosomes. PubMed DOI PMC

LaCount D. J., Barrett B. and Donelson J. E. (2002). PubMed DOI

Logan D. C., Scott I. and Tobin A. K. (2003). The genetic control of plant mitochondrial morphology and dynamics. PubMed DOI

Matthews K. R. (2011). Controlling and coordinating development in vector-transmitted parasites. PubMed DOI PMC

McCulloch R., Vassella E., Burton P., Boshart M. and Barry J. D. (2004). Transformation of monomorphic and pleomorphic PubMed DOI

Morriswood B., Havlicek K., Demmel L., Yavuz S., Sealey-Cardona M., Vidilaseris K., Anrather D., Kostan J., Djinovic-Carugo K., Roux K. J. et al. (2013). Novel bilobe components in PubMed DOI PMC

Nozaki T., Haynes P. A. and Cross G. A. M. (1996). Characterization of the PubMed DOI

Ogbadoyi E. O., Robinson D. R. and Gull K. (2003). A high-order trans-membrane structural linkage is responsible for mitochondrial genome positioning and segregation by flagellar basal bodies in trypanosomes. PubMed DOI PMC

Ooi C.-P. and Bastin P. (2013). More than meets the eye: understanding PubMed DOI PMC

Poon S. K., Peacock L., Gibson W., Gull K. and Kelly S. (2012). A modular and optimized single marker system for generating PubMed DOI PMC

Price H. P., Hodgkinson M. R., Curwen R. S., MacLean L. M., Brannigan J. A., Carrington M., Smith B. A., Ashford D. A., Stark M. and Smith D. F. (2012). The orthologue of Sjögren's syndrome nuclear autoantigen 1 (SSNA1) in PubMed DOI PMC

Reynolds E. S. (1963). The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. PubMed DOI PMC

Robinson D. R. and Gull K. (1991). Basal body movements as a mechanism for mitochondrial genome segregation in the trypanosome cell cycle. PubMed DOI

Robinson D. R., Sherwin T., Ploubidou A., Byard E. H. and Gull K. (1995). Microtubule polarity and dynamics in the control of organelle positioning, segregation, and cytokinesis in the trypanosome cell cycle. PubMed DOI PMC

Rotureau B., Blisnick T., Subota I., Julkowska D., Cayet N., Perrot S. and Bastin P. (2014). Flagellar adhesion in PubMed DOI

Schimanski B., Nguyen T. N. and Günzl A. (2005). Highly efficient tandem affinity purification of trypanosome protein complexes based on a novel epitope combination. PubMed DOI PMC

Schneider C. A., Rasband W. S. and Eliceiri K. W. (2012). NIH Image to ImageJ: 25 years of image analysis. PubMed DOI PMC

Sen A., Damm V. T. and Cox R. T. (2013). PubMed DOI PMC

Sharma R., Gluenz E., Peacock L., Gibson W., Gull K. and Carrington M. (2009). The heart of darkness: growth and form of PubMed DOI PMC

Sherwin T. and Gull K. (1989). The cell division cycle of PubMed DOI

Sun S. Y., Wang C., Yuan Y. A. and He C. Y. (2013). An intracellular membrane junction consisting of flagellum adhesion glycoproteins links flagellum biogenesis to cell morphogenesis in PubMed DOI

Sunter J. D., Varga V., Dean S. and Gull K. (2015). A dynamic coordination of flagellum and cytoplasmic cytoskeleton assembly specifies cell morphogenesis in trypanosomes. PubMed DOI PMC

Urwyler S., Studer E., Renggli C. K. and Roditi I. (2007). A family of stage-specific alanine-rich proteins on the surface of epimastigote forms of PubMed DOI

Vaughan S., Kohl L., Ngai I., Wheeler R. J. and Gull K. (2008). A repetitive protein essential for the flagellum attachment zone filament structure and function in PubMed DOI

Verner Z., Basu S., Benz C., Dixit S., Dobáková E., Faktorová D., Hashimi H., Horáková E., Huang Z., Paris Z. et al. (2015). Malleable mitochondrion of PubMed DOI

Wheeler R. J., Scheumann N., Wickstead B., Gull K. and Vaughan S. (2013). Cytokinesis in PubMed DOI PMC

Wickstead B., Ersfeld K. and Gull K. (2002). Targeting of a tetracycline-inducible expression system to the transcriptionally silent minichromosomes of PubMed DOI

Zhou Q., Liu B., Sun Y. and He C. Y. (2011). A coiled-coil- and C2-domain-containing protein is required for FAZ assembly and cell morphology in PubMed DOI

Zhou Q., Hu H., He C. Y. and Li Z. (2015). Assembly and maintenance of the flagellum attachment zone filament in PubMed DOI PMC

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