A novel method to analyze nucleocytoplasmic transport in vivo by using short peptide tags.
J Mol Biol
; 425(10): 1839-45, 2013 May 27.
Article
em En
| MEDLINE
| ID: mdl-23416199
ABSTRACT
Regulated nucleocytoplasmic transport is of vital importance for maintaining the physiology of the cell, and disturbed nucleocytoplasmic shuttling of certain proteins has been found in a variety of diseases including cancer. The most frequently used procedure to analyze those processes is to fuse the protein of interest to a fluorescent protein such as GFP (green fluorescent protein)--a technique that is prone to impair normal protein function and subcellular localization. We report a novel approach to monitor nucleocytoplasmic transport processes in vivo by combining short TetR inducing peptide tags (TIP) with a TetR-controlled reporter gene in a human cell line. The technology is exemplified by demonstrating nucleocytoplasmic shuttling of the glucocorticoid receptor and activity of two further TIP fusions to cancer-related proteins. The technology presented provides the basis for efficient screening systems to isolate compounds altering the nucleocytoplasmic distribution of a protein of interest.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Transporte Ativo do Núcleo Celular
Limite:
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article