查看文章

Polo-like kinases (Plks) play crucial roles in mitosis and cell division. Whereas lower eukaryotes typically contain a single Plk, mammalian cells express several closely related but functionally distinct Plks. We describe here a chemical genetic system in which a single Plk family member, Plk1, can be inactivated with high selectivity and temporal resolution by using an allele-specific, small-molecule inhibitor, as well as the application of this system to dissect Plk1's role in cytokinesis. To do this, we disrupted both copies of the PLK1 locus in human cells through homologous recombination and then reconstituted Plk1 activity by using either the wild-type kinase (Plk1^wt) or a mutant version whose catalytic pocket has been enlarged to accommodate bulky purine analogs (Plk1^as). When cultured in the presence of these analogs, Plk1^as cells accumulate in prometaphase with defects that parallel those found in PLK1^Δ/Δ …