Trypanosoma brucei brucei: endocytic recycling is important for mouse infectivity.

Journal article

Endocytosis in the African trypanosome, Trypanosoma brucei, is intimately involved in maintaining homeostasis of the cell surface proteome, morphology of the flagellar pocket and has recently been demonstrated as a bona fide drug target. RNAi-mediated knockdown of many factors required for endocytic transport, including several small GTPases, the major coat protein clathrin and a clathrin-associated receptor, epsinR, results in rapid cell death in vitro. Rapid loss of viability in vitro precludes meaningful investigation by RNAi of the roles of trypanosome endocytosis in vivo. Here we have sought to address this issue using strategies designed to produce milder effects on the endocytic system than complete functional ablation. We created a trypanosome clathrin heavy chain hemizygote and several lines expressing mutant forms of Rab5 and Rab11, described previously. All are viable in in vitro culture, with negligible impact to proliferative rates or cell cycle. Clathrin hemizygotes express clathrin heavy chain at ∼50% of wild type levels, but despite this demonstrate no defect to growth in mice, while none of the Rab5 mutants affected proliferation in vivo, despite clear evidence for effects on endocytosis. By contrast we find that expressing a dominantly active Rab11 mutant led to compromised growth in mice. These data indicate that trypanosomes likely tolerate the effects of partly decreased clathrin expression and alterations in early endocytosis, but are more sensitive to alterations in the recycling arm of the pathway.

Authors: Natesan, S; Black, A; Matthews, K; Mottram, J; Field, M

• Journal: Experimental parasitology
• Volume: 127
• Issue: 4
• Pages: 777-783
• Publication date: 2011-04-01
• DOI: 10.1016/j.exppara.2011.01.001
• EISSN: 1090-2449
• ISSN: 0014-4894
• Language: English
• Keywords: rab5 GTP-Binding Proteins; Endocytosis; Trypanosomiasis, African; Animals; Trypanosoma brucei brucei; Gene Knockout Techniques; rab GTP-Binding Proteins; Mice; Disease Models, Animal; Mice, Inbred ICR; Gene Expression Regulation; Clathrin Heavy Chains; Parasitemia