Clathrin expression in Trypanosoma cruzi

Ligia Cristina Kalb,Iriane Eger,Stênio Perdigão Fragoso,Cassiano Martin Batista,Maurilio José Soares,Yohana Camila Antunes Frederico

doi:10.1186/1471-2121-15-23

Ligia Cristina Kalb, Iriane Eger + Show 4 more

Open Access

https://doi.org/10.1186/1471-2121-15-23

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Abstract

BackgroundClathrin-mediated vesicular trafficking, the mechanism by which proteins and lipids are transported between membrane-bound organelles, accounts for a large proportion of import from the plasma membrane (endocytosis) and transport from the trans-Golgi network towards the endosomal system. Clathrin-mediated events are still poorly understood in the protozoan Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. In this study, clathrin heavy (TcCHC) and light (TcCLC) chain gene expression and protein localization were investigated in different developmental forms of T. cruzi (epimastigotes, trypomastigotes and amastigotes), using both polyclonal and monoclonal antibodies raised against T. cruzi recombinant proteins.ResultsAnalysis by confocal microscopy revealed an accumulation of TcCHC and TcCLC at the cell anterior, where the flagellar pocket and Golgi complex are located. TcCLC partially colocalized with the Golgi marker TcRAB7-GFP and with ingested albumin, but did not colocalize with transferrin, a protein mostly ingested via uncoated vesicles at the cytostome/cytopharynx complex.ConclusionClathrin heavy and light chains are expressed in T. cruzi. Both proteins typically localize anterior to the kinetoplast, at the flagellar pocket and Golgi complex region. Our data also indicate that in T. cruzi epimastigotes clathrin-mediated endocytosis of albumin occurs at the flagellar pocket, while clathrin-independent endocytosis of transferrin occurs at the cytostome/cytopharynx complex.

Highlights

Clathrin-mediated vesicular trafficking, the mechanism by which proteins and lipids are transported between membrane-bound organelles, accounts for a large proportion of import from the plasma membrane and transport from the trans-Golgi network towards the endosomal system
TcCLC and TcCHC: in silico gene identification and domain analysis A hypothetical gene for T. cruzi clathrin light chain was identified in the TritrypDB database (Tc00.1047053506211.240)
The T. cruzi clathrin heavy chain gene has already been identified in the TritrypDB database

Summary

Introduction

Clathrin-mediated vesicular trafficking, the mechanism by which proteins and lipids are transported between membrane-bound organelles, accounts for a large proportion of import from the plasma membrane (endocytosis) and transport from the trans-Golgi network towards the endosomal system. The most thoroughly characterized form of clathrin comprises light chain subunits, which are unstructured until bound to heavy chains [1]. Clathrin-mediated vesicular trafficking is the most highly characterized mechanism by which proteins and lipids are transported between membrane-bound organelles, being responsible for a large proportion of import from the plasma membrane (endocytosis) and transport from the trans-Golgi network (TGN) towards the endosomal system [4,5]. A good example of these parasitic protozoa are trypanosomatids, a group that includes pathogenic parasites, such as Trypanosoma brucei, T. cruzi and different Leishmania species, which are all associated with severe diseases in humans

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