Green fluorescent protein tag for studies of drug-induced translocation of nucleolar protein RH-II/Gu

被引：16

作者：

Valdez, BC ^{[1
]}

Perlaky, L ^{[1
]}

Cai, ZJ ^{[1
]}

Henning, D ^{[1
]}

Busch, H ^{[1
]}

机构：

[1] Baylor Coll Med, Dept Pharmacol, Houston, TX 77030 USA

来源：

BIOTECHNIQUES | 1998年 / 24卷 / 06期

关键词：

D O I：

10.2144/98246cr03

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We have constructed a human osteogenic sarcoma cell line, U-2 OS/GFP-Gu, that expresses nucleolar RNA helicase RH-II/Gu tagged with green fluorescent protein (GFP). The presence of a GFP tag does nor inhibit RNA helicase. RNA folding and ATPase activities of RH-II/Gu protein. The derived cell line responds to cytotoxic agents like the parental cell line U-2 OS. In the presence of either actinomycin D or toyocamycin, the GFP-RH-II/Gu fusion protein translocates from the nucleolus to the nucleoplasm in the same way as the translocation of endogenous RH-II/Gu. The drug-induced translocation of GFP-RH-II/Gu is easily monitored by direct observation of live cells in vivo. This cell line can be used to screen cytotoxic drugs and to study the mechanisms of drug-induced translocation of RH-II/Gu. The cellular localization of RH-II/Gu during the cell cycle-dependent formation of the nucleolus is readily monitored. Real-time results are obtained more quickly without disadvantages associated with cell fixation and immunofluorescence-based staining.

引用

页码：1032 / 1036

页数：5

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