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Journal of Biomedicine and Biotechnology
Volume 2012 (2012), Article ID 657942, 15 pages
http://dx.doi.org/10.1155/2012/657942
Research Article

EhADH112 Is a Bro1 Domain-Containing Protein Involved in the Entamoeba histolytica Multivesicular Bodies Pathway

1Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, 07360 México, DF, Mexico
2Centro de Diagnóstico y Vigilancia Epidemiológica del Distrito Federal, Instituto de Ciencia y Tecnología del Distrito Federal, 06010 México, DF, Mexico
3Escuela Superior de Medicina, Instituto Politécnico Nacional, 11340 México, DF, Mexico
4Instituto de Investigaciones Biomédicas “Alberto Sols”, CSIC-UAM, 28029 Madrid, Spain
5Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, 03100 México, DF, Mexico

Received 16 July 2011; Accepted 3 October 2011

Academic Editor: Luis I. Terrazas

Copyright © 2012 Cecilia Bañuelos et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

EhADH112 is an Entamoeba histolytica Bro1 domain-containing protein, structurally related to mammalian ALIX and yeast BRO1, both involved in the Endosomal Sorting Complexes Required for Transport (ESCRT)-mediated multivesicular bodies (MVB) biogenesis. Here, we investigated an alternative role for EhADH112 in the MVB protein trafficking pathway by overexpressing 166 amino acids of its N-terminal Bro1 domain in trophozoites. Trophozoites displayed diminished phagocytosis rates and accumulated exogenous Bro1 at cytoplasmic vesicles which aggregated into aberrant complexes at late stages of phagocytosis, probably preventing EhADH112 function. Additionally, the existence of a putative E. histolytica ESCRT-III subunit (EhVps32) presumably interacting with EhADH112, led us to perform pull-down experiments with GST-EhVps32 and [35S]-labeled EhADH112 or EhADH112 derivatives, confirming EhVps32 binding to EhADH112 through its Bro1 domain. Our overall results define EhADH112 as a novel member of ESCRT-accessory proteins transiently present at cellular surface and endosomal compartments, probably contributing to MVB formation during phagocytosis.