FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins

被引：12

作者：

Belaya, K ^{[1
]}

Tollervey, D ^{[1
]}

Kos, M ^{[1
]}

机构：

[1] Univ Edinburgh, Wellcome Trust Ctr Cell Biol, Edinburgh EH9 3JR, Midlothian, Scotland

来源：

RNA | 2006年 / 12卷 / 05期

基金：

英国惠康基金;

关键词：

nucleo-cytoplasmic shuttling; Saccharomyces cerevisiae;

D O I：

10.1261/rna.2301806

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：921 / 924

页数：4

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