A novel method to analyze nucleocytoplasmic transport in vivo by using short peptide tags.

Cornelius Wimmer, Stephanie Platzer, Wolfgang Hillen, Marcus Klotzsche

Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, 91058 Erlangen, Germany.

Journal of molecular biology 2013 May 27

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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. Copyright © 2013 Elsevier Ltd. All rights reserved.

Citation

Cornelius Wimmer, Stephanie Platzer, Wolfgang Hillen, Marcus Klotzsche. A novel method to analyze nucleocytoplasmic transport in vivo by using short peptide tags. Journal of molecular biology. 2013 May 27;425(10):1839-45