High-content classification of nucleocytoplasmic import or export inhibitors.
Transcription factors of the nuclear factor kappa 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 EC(50) 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 IkappaB 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 kappa B.
Kwon
Yong-Jun
YJ
Institut Pasteur-Korea, Seongbuk-Gu, Seoul, Korea.
Genovesio
Auguste
A
Youl Kim
Nam
N
Hi Chul Kim
Jung
Sungyong
S
David-Watine
Brigitte
B
Nehrbass
Ulf
U
Emans
Neil
N
D021581:Active Transport, Cell Nucleus / Q000187:drug effects*; D000465:Algorithms; D000880:Anthraquinones / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects*; D003198:Computer Simulation; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D020672:I-kappa B Proteins / Q000037:antagonists & inhibitors; D020128:Inhibitory Concentration 50; D007668:Kidney / Q000166:cytology; D008954:Models, Biological; D016328:NF-kappa B / Q000096:biosynthesis; D009570:Nitriles / Q000494:pharmacology; D010703:Phorbol Esters / Q000494:pharmacology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D013450:Sulfones / Q000494:pharmacology; D013997:Time Factors; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
C520537:1,5-bis((2-(methylamino)ethyl)amino)-4,8-dihydroxyanthracene-9,10-dione; C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D000880:Anthraquinones; D004791:Enzyme Inhibitors; D005231:Fatty Acids, Unsaturated; D005419:Flavonoids; C083487:I kappa B beta protein; D020672:I-kappa B Proteins; D016328:NF-kappa B; D009570:Nitriles; D010703:Phorbol Esters; D013450:Sulfones; D048051:p38 Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; C038753:leptomycin B