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Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of proteomic data available shows that many of these processes are highly dynamic. Therefore, methods are needed to reliably assess whether a protein shuttles between nucleus and cytoplasm, and the kinetics with which it exchanges. Here we describe a combination of the classical heterokaryon assay with fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) techniques, which allows an assessment of the kinetics of protein shuttling in the yeast Saccharomyces cerevisiae.

Original publication

DOI

10.1261/rna.2301806

Type

Journal article

Journal

Rna

Publication Date

05/2006

Volume

12

Pages

921 - 930

Keywords

Active Transport, Cell Nucleus, Cell Nucleus, Cytoplasm, Fluorescence Recovery After Photobleaching, Kinetics, Microscopy, Fluorescence, Plasmids, Protein Transport, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins