Inducible translocation trap: a system for detecting inducible nuclear translocation

Molecular Cell
Akemi HoshinoHodaka Fujii

Abstract

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.

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Citations

Dec 22, 2009·The Journal of Biological Chemistry·Christopher N ParkhurstMoses V Chao
Aug 11, 2007·Cell Stress & Chaperones·Akemi Hoshino, Hodaka Fujii
Oct 30, 2008·Journal of Immunotoxicology·Hodaka Fujii
Feb 3, 2007·Biochemical and Biophysical Research Communications·Hodaka Fujii
Jun 12, 2008·Developmental Biology·Alexander Emelyanov, Serguei Parinov
Apr 21, 2007·The Journal of Biological Chemistry·Akemi HoshinoHodaka Fujii

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