A functional study of nucleocytoplasmic transport signals of the EhNCABP166 protein from Entamoeba histolytica
Por:
Uribe R., Ma, JDB, Robles-Flores M., Hernandez, GM, González-Robles A., Hernández-Rivas R., Guillen N., Vargas M.
Publicada:
1 nov 2012
Resumen:
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 domai
Filiaciones:
Uribe R.:
Univ Nacl Autonoma Mexico, Programa Doctorado Ciencias Biomed, Mexico City 04510, DF, Mexico
Robles-Flores M.:
Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Mexico City 04510, DF, Mexico
Hernandez, GM:
Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Mexico City 04510, DF, Mexico
González-Robles A.:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico
Hernández-Rivas R.:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico
Guillen N.:
Institut Pasteur, Unité Biologie Cellulaire du Parasitisme, Paris, F-75015, France
Vargas M.:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico
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