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Eukaryotic Cell, May 2005, p. 971-980, Vol. 4, No. 5
1535-9778/05/$08.00+0     doi:10.1128/EC.4.5.971-980.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Developmental Variation in Rab11-Dependent Trafficking in Trypanosoma brucei

Belinda S. Hall, Emma Smith, Wolfram Langer, Louisa A. Jacobs, David Goulding, and Mark C. Field*

Department of Biological Sciences, Imperial College, Exhibition Road, London SW7 2AY, United Kingdom

Received 7 December 2004/ Accepted 16 March 2005

In Trypanosoma brucei, endocytosis is developmentally regulated and is substantially more active in the mammalian infective stage, where it likely plays a role in immune evasion. The small GTPase TbRAB11 is highly expressed in the mammalian stage and mediates recycling of glycosylphosphatidylinositol-anchored proteins, including the variant surface glycoprotein (VSG) and the transferrin receptor, plus trafficking of internalized anti-VSG antibody and transferrin. No function has been assigned to TbRAB11 in the procyclic (insect) stage trypanosome. The importance of TbRAB11 to both bloodstream and procyclic form viability was assessed by RNA interference (RNAi). Suppression of TbRAB11 in the bloodstream form was rapidly lethal and led to cells with round morphology and an enlarged flagellar pocket. TbRAB11 RNAi was also lethal in procyclic forms, which also became rounded, but progression to cell death was significantly slower and the flagellar pocket remained normal. In bloodstream forms, silencing of TbRAB11 had no effect on exocytosis of newly synthesized VSG, fluid-phase endocytosis, or transferrin uptake, but export of internalized transferrin was inhibited. Lectin endocytosis assays revealed a block to postendosomal transport mediated by suppressing TbRAB11. By contrast, in procyclic forms, depletion of TbRAB11 blocks both fluid-phase endocytosis and internalization of surface proteins. In normal bloodstream forms, most VSG is recycled, but in procyclics, internalized surface proteins accumulated in the lysosome. These data demonstrate that TbRAB11 controls recycling and is essential in both life stages of T. brucei but that its primary role is subject to developmental variation.


* Corresponding author. Present address: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, United Kingdom. Phone: 44 01223 333734. Fax: 44 01223 333734. E-mail: mcf34{at}cam.ac.uk.


Eukaryotic Cell, May 2005, p. 971-980, Vol. 4, No. 5
1535-9778/05/$08.00+0     doi:10.1128/EC.4.5.971-980.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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