High-Content Classification of Nucleocytoplasmic Import or Export Inhibitors
Yong-Jun Kwon,
Auguste Genovesio,
Nam Youl Kim,
Hi Chul Kim,
Sungyong Jung,
Brigitte David-Watine,
Ulf Nehrbass,
Neil Emans*
Institut Pasteur Korea
* To whom correspondence should be addressed. E-mail: emans{at}pasteur.or.kr.
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Abstract |
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Transcription factors of the nuclear factor 
B family are the paradigm for signaling dependent nuclear translocation and are ideally suited to analysis through image-based chemical genetic screening. The authors describe combining high-content image analysis with a compound screen to identify compounds affecting either nuclear import or export. Validation in silico and in vitro determined an EC50 for the nuclear export blocker leptomycin B of 2.4 ng/mL (4.4 nM). The method demonstrated high selectivity (Z'>0.95), speed, and robustness in a screen of a compound collection. It identified the IB protein kinase inhibitor BAY 11 7082 as an import inhibitor, the p38 mitogen-activated protein (MAP) kinase inhibitor PD98509 as an import enhancer, and phorbol ester as an export inhibitor. The results establish a robust method for identifying compounds regulating nucleocytoplasmic import or export and also implicate MAP kinases in nuclear import of nuclear factor B. (Journal of Biomolecular Screening XXXX:xx-xx)
