Cell-permeable small molecules that inhibit their targets on fast timescales are powerful probes of cell-division mechanisms. Such inhibitors have been identified using phenotype-based screens with chemical libraries. However, the characteristics of compound libraries needed to effectively span cell-division phenotype space, to find probes that target different mechanisms, are not known. Here we show that a small collection of 100 diaminopyrimidines (DAPs) yields a range of cell-division phenotypes, including changes in spindle geometry, chromosome positioning and mitotic index. Monopolar mitotic spindles are induced by four inhibitors, including one that targets Polo-like kinases (Plks), evolutionarily conserved serine/threonine kinases. Using chemical inhibitors and high-resolution live-cell microscopy, we found that Plk activity is needed for the assembly and maintenance of bipolar mitotic spindles. Plk inhibition destabilizes kinetochore microtubules while stabilizing other spindle microtubules, leading to monopolar spindles. Further testing of compounds based on 'privileged scaffolds', such as the DAP scaffold, could lead to new cell-division probes and antimitotic agents.
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We thank C. Karan and acknowledge the use of the Rockefeller University High Throughput Screening Resource Center. This work was supported by US National Institutes of Health grant GM71772 (to T.M.K.).
The authors declare no competing financial interests.
Composition of the diaminopyrimidine library. (PDF 194 kb)
Overview of cell-based screening results. (PDF 257 kb)
Mitotic phenotypes induced by a subset of DAPs. (PDF 442 kb)
Monopolar mitotic spindles induced by two diaminopyrimidines do not result from changes in centrosome number or Eg5 inhibition. (PDF 908 kb)
BTO-1 inhibits Polo-like kinase 1 in vitro. (PDF 93 kb)
Additional images for comparing Plk1-knock-down and treatment with chemical inhibitors. (PDF 129 kb)
Quantitation of monopolar spindles and mitotic indices in DAP-81 and BTO-1 treated PTKαT cells. (PDF 97 kb)
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