Inducible translocation trap: A system for detecting inducible nuclear translocation

被引：20

作者：

Hoshino, A ^{[1
]}

Matsumura, S ^{[1
]}

Kondo, K ^{[1
]}

Hirst, JA ^{[1
]}

Fujii, H ^{[1
]}

机构：

[1] NYU, Sch Med, Dept Pathol, New York, NY 10016 USA

来源：

MOLECULAR CELL | 2004年 / 15卷 / 01期

关键词：

D O I：

10.1016/j.molcel.2004.06.017

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

Here we report a general system, inducible translocation trap (ITT), for identification of proteins that translocate into the nucleus following signal transduction from cell surface receptors. ITT consists of a retroviral cDNA expression library of fusion proteins consisting of a LexA DNA binding domain, the transactivation domain of a transcriptional activator, and proteins encoded by cDNA inserts. The retroviral library is then transduced into cell lines containing a reporter gene with LexA binding sites in its promoter. Cells expressing the reporter gene by extracellular stimuli are then selected by flow-cytometric sorting. By using ITT, we identified cDNA encoding Stat1 in a screen of proteins which translocate into the nucleus by IFNgamma, indicating that this system can be used for isolation of nuclear translocating proteins induced by extracellular stimuli. ITT may be a useful tool for dissecting dynamic translocation in various biological systems.

引用

页码：153 / 159

页数：7

共 33 条

[1]

Two co-existing mechanisms for nuclear import of MAP kinase: passive diffusion of a monomer and active transport of a dimer [J].