A Protein-Protein Interaction Map of the Trypanosoma brucei Paraflagellar Rod

Adam Yuan,Sylvain Lacomble,Neil Portman,Keith Gull

doi:10.1371/journal.pone.0007685

Abstract

We have conducted a protein interaction study of components within a specific sub-compartment of a eukaryotic flagellum. The trypanosome flagellum contains a para-crystalline extra-axonemal structure termed the paraflagellar rod (PFR) with around forty identified components. We have used a Gateway cloning approach coupled with yeast two-hybrid, RNAi and 2D DiGE to define a protein-protein interaction network taking place in this structure. We define two clusters of interactions; the first being characterised by two proteins with a shared domain which is not sufficient for maintaining the interaction. The other cohort is populated by eight proteins, a number of which possess a PFR domain and sub-populations of this network exhibit dependency relationships. Finally, we provide clues as to the structural organisation of the PFR at the molecular level. This multi-strand approach shows that protein interactome data can be generated for insoluble protein complexes.

Highlights

Trypanosomatid protozoan parasites are the causative agents of a number of diseases responsible for the death of thousands of people in developing countries
All trypanosomes produce a single flagellum which is involved in numerous aspects of parasite biology including motility, cytokinesis, environment sensing, attachment to the host [1,2,3,4,5,6] and in the case of the African trypanosome Trypanosoma brucei, correct generation and function of the flagellum is essential for the survival of the mammalian bloodstream stage [7]
8 nonredundant interactions were detected in the yeast two-hybrid screen (Figure 1) and these can be divided into two clusters: PAR1PFC3-PFR5-PFC20-PFC6-PFR6 and PFC4-PFC16

Summary

Introduction

Trypanosomatid protozoan parasites are the causative agents of a number of diseases responsible for the death of thousands of people in developing countries. All trypanosomes produce a single flagellum which is involved in numerous aspects of parasite biology including motility, cytokinesis, environment sensing, attachment to the host [1,2,3,4,5,6] and in the case of the African trypanosome Trypanosoma brucei, correct generation and function of the flagellum is essential for the survival of the mammalian bloodstream stage [7]. The flagellum incorporates the canonical 9+2 microtubular eukaryotic axoneme and, as with the flagella of many species, has several additional lineage specific features. One of these additional features is an extraaxonemal para-crystalline structure termed the paraflagellar rod (PFR). In addition to its role in motility [10,11], the PFR serves as a platform for metabolic and signaling enzymes [2,12,13] and is essential for the survival of the mammalian infective form of the parasite in the host bloodstream [14]

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Conclusion