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High‐Content Screening and Profiling of Drug Activity in an Automated Centrosome‐Duplication Assay
Author(s) -
Perlman Zachary E.,
Mitchison Timothy J.,
Mayer Thomas U.
Publication year - 2005
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200400266
Subject(s) - centrosome , high content screening , gene duplication , biology , microtubule , computational biology , microbiology and biotechnology , retinoic acid , cell cycle , gene , cell , genetics
Maintenance of centrosome number is essential for cell‐cycle progression and genomic stability, but investigation of this regulation has been limited by assay difficulty. We present a fully automated image‐based centrosome‐duplication assay that is accurate and robust enough for both careful cell‐biology studies and high‐throughput screening, and employ this assay in a series of chemical‐genetic studies. We observe that a simple cytometric profiling strategy, which is based on organelle size, groups compounds with similar mechanisms of action; this suggests a simple strategy for excluding compounds that undesirably target such activities as protein synthesis and microtubule dynamics. Screening a library of compounds of known activity, we found unexpected effects on centrosome duplication by a number of drugs, most notably isoform‐specific protein kinase C inhibitors and retinoic acid receptor agonists. From a 16 320‐member library of uncharacterized small molecules, we identified five potent centrosome‐duplication inhibitors that do not target microtubule dynamics or protein synthesis. The analysis methodology reported here is directly relevant to studies of centrosome regulation in a variety of systems and is adaptable to a wide range of other biological problems.