Green Fluorescent Protein Tag for Studies of Drug-Induced Translocation of Nucleolar Protein RH-II/Gu | Zendy

Benigno C. Valdez | Zendy; László Perlaky | Zendy; Zhengguo Cai | Zendy; D Henning | Zendy; H. Busch | Zendy

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Green Fluorescent Protein Tag for Studies of Drug-Induced Translocation of Nucleolar Protein RH-II/Gu

Author(s) -

Benigno C. Valdez,

László Perlaky,

Zhengguo Cai,

D Henning,

H. Busch

Publication year - 1998

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/98246cr03

Subject(s) - nucleolus , green fluorescent protein , nucleoplasm , microbiology and biotechnology , helicase , chromosomal translocation , biology , fusion protein , cell culture , rna , cell cycle , cell , chemistry , cytoplasm , biochemistry , gene , recombinant dna , genetics

We have constructed a human osteogenic sarcoma cell line, U-2 OS/GFPGu, that expresses nucleolar RNA helicase RH-II/Gu tagged with green fluorescent protein (GFP). The presence of a GFP tag does not 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 druginduced 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 the disadvantages associated with cell fixation and immunofluorescence-based staining.

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