FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins

K Belaya, D Tollervey, M Kos

Research output: Contribution to journalArticlepeer-review

Abstract

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 languageEnglish
Pages (from-to)921-924
Number of pages4
JournalRNA
Volume12
Issue number5
DOIs
Publication statusPublished - May 2006

Keywords

  • nucleo-cytoplasmic shuttling
  • Saccharomyces cerevisiae

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