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A functional study of nucleocytoplasmic transport signals of the EhNCABP166 protein from Entamoeba histolytica

Published online by Cambridge University Press:  23 August 2012

R. URIBE ,
J. ALMARAZ BARRERA MA DE ,
M. ROBLES-FLORES ,
G. MENDOZA HERNÁNDEZ ,
A. GONZÁLEZ-ROBLES ,
R. HERNÁNDEZ-RIVAS ,
N. GUILLEN  and
M. VARGAS
R. URIBE
Affiliation:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F, México Programa de Doctorado en Ciencias Biomédicas, Universidad Nacional Autónoma de México, D.F, México
J. ALMARAZ BARRERA MA DE
Affiliation:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F, México
M. ROBLES-FLORES
Affiliation:
Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, D.F, México
G. MENDOZA HERNÁNDEZ
Affiliation:
Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, D.F, México
A. GONZÁLEZ-ROBLES
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F, México
R. HERNÁNDEZ-RIVAS
Affiliation:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F, México
N. GUILLEN
Affiliation:
Institut Pasteur, Unité Biologie Cellulaire du Parasitisme, Paris, F-75015, France
M. VARGAS*
Affiliation:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F, México
*
*Corresponding author: Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F, México. Tel: +52 55 57473326. Fax: +52 55 5747 3326. E-mail address: [email protected]
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Summary

EhNCABP166 is an Entamoeba histolytica actin-binding protein that localizes to the nucleus and cytoplasm. Bioinformatic analysis of the EhNCABP166 amino acid sequence shows the presence of 3 bipartite nuclear localization signals (NLS) and a nuclear export signal (NES). The present study aimed to investigate the functionality of these signals in 3 ways. First, we fused each potential NLS to a cytoplasmic domain of ehFLN to determine whether the localization of this domain could be altered by the presence of the NLSs. Furthermore, the localization of each domain of EhNCABP166 was determined. Similarly, we generated mutations in the first block of bipartite signals from the domains that contained these signals. Additionally, we added an NES to 2 constructs that were then evaluated. We confirmed the intranuclear localization of EhNCABP166 using transmission electron microscopy. Fusion of each NLS resulted in shuttling of the cytoplasmic domain to the nucleus. With the exception of 2 domains, all of the evaluated domains localized within the nucleus. A mutation in the first block of bipartite signals affected the localization of the domains containing an NLS. The addition of an NES shifted the localization of these domains to the cytoplasm. The results presented here establish EhNCABP166 as a protein containing functional nuclear localization signals and a nuclear export signal.

Keywords

Entamoeba histolyticanuclear localization signal (NLS)nuclear export signal (NES)actin-binding proteinnucleocytoplasmic transport system
Type
Research Article
Information
Parasitology , Volume 139 , Issue 13 , November 2012 , pp. 1697 - 1710
Copyright
Copyright © Cambridge University Press 2012

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