2010年4月
Rapid-Response Splicing Reporter Screens Identify Differential Regulators of Constitutive and Alternative Splicing
MOLECULAR AND CELLULAR BIOLOGY
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- 巻
- 30
- 号
- 7
- 開始ページ
- 1718
- 終了ページ
- 1728
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1128/MCB.01301-09
- 出版者・発行元
- AMER SOC MICROBIOLOGY
Bioactive compounds have been invaluable for dissecting the mechanisms, regulation, and functions of cellular processes. However, very few such reagents have been described for pre-mRNA splicing. To facilitate their systematic discovery, we developed a high-throughput cell-based assay that measures pre-mRNA splicing by utilizing a quantitative reporter system with advantageous features. The reporter, consisting of a destabilized, intron-containing luciferase expressed from a short-lived mRNA, allows rapid screens (<4 h), thereby obviating the potential toxicity of splicing inhibitors. We describe three inhibitors (out of >23,000 screened), all pharmacologically active: clotrimazole, flunarizine, and chlorhexidine. Interestingly, none was a general splicing inhibitor. Rather, each caused distinct splicing changes of numerous genes. We further discovered the target of action of chlorhexidine and show that it is a selective inhibitor of specific Cdc2-like kinases (Clks) that phosphorylate serine-arginine-rich (SR) protein splicing factors. Our findings reveal unexpected activities of clinically used drugs in splicing and uncover differential regulation of constitutively spliced introns.
- リンク情報
- ID情報
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- DOI : 10.1128/MCB.01301-09
- ISSN : 0270-7306
- eISSN : 1098-5549
- PubMed ID : 20123975
- Web of Science ID : WOS:000275302000012