title: Stathmin and microtubules regulate mitotic entry in HeLa cells by controlling activation of both Aurora kinase A and Plk1.
abstract: Depletion of stathmin, a microtubule (MT) destabilizer, delays mitotic entry by ∼4 h in HeLa cells. Stathmin depletion reduced the activity of CDC25 and its upstream activators, Aurora A and Plk1. Chemical inhibition of both Aurora A and Plk1 was sufficient to delay mitotic entry by 4 h, while inhibiting either kinase alone did not cause a delay. Aurora A and Plk1 are likely regulated downstream of stathmin, because the combination of stathmin knockdown and inhibition of Aurora A and Plk1 was not additive and again delayed mitotic entry by 4 h. Aurora A localization to the centrosome required MTs, while stathmin depletion spread its localization beyond that of γ-tubulin, indicating an MT-dependent regulation of Aurora A activation. Plk1 was inhibited by excess stathmin, detected in in vitro assays and cells overexpressing stathmin-cyan fluorescent protein. Recruitment of Plk1 to the centrosome was delayed in stathmin-depleted cells, independent of MTs. It has been shown that depolymerizing MTs with nocodazole abrogates the stathmin-depletion induced cell cycle delay; in this study, depolymerization with nocodazole restored Plk1 activity to near normal levels, demonstrating that MTs also contribute to Plk1 activation. These data demonstrate that stathmin regulates mitotic entry, partially via MTs, to control localization and activation of both Aurora A and Plk1.
authors.affiliation: Department of Biological Sciences, Lehigh University, Bethlehem, PA 18015.
authors.forename: Victoria C
authors.initials: VC
authors.lastname: Silva
authors.forename: Lynne
authors.initials: L
authors.lastname: Cassimeris
mesh_terms: D064096:Aurora Kinase A / Q000037:antagonists & inhibitors*; D016203:CDC2 Protein Kinase / Q000235:genetics; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors*; D045744:Cell Line, Tumor; D018385:Centrosome; D019926:Cyclin B / Q000235:genetics; D059565:G2 Phase Cell Cycle Checkpoints / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008870:Microtubules / Q000235:genetics*; D008938:Mitosis / Q000187:drug effects*; D015739:Nocodazole / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors*; D011621:Pteridines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering; D050777:Stathmin / Q000235:genetics*; D014404:Tubulin; D050257:Tubulin Modulators / Q000494:pharmacology; D020687:cdc25 Phosphatases; D000097869:Polo-Like Kinase 1
chemical_list: D064096:Aurora Kinase A; D016203:CDC2 Protein Kinase; D018797:Cell Cycle Proteins; D019926:Cyclin B; D015739:Nocodazole; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D011621:Pteridines; D034741:RNA, Small Interfering; D050777:Stathmin; D014404:Tubulin; D050257:Tubulin Modulators; D020687:cdc25 Phosphatases; D000097869:Polo-Like Kinase 1; C518477:BI 2536; C576917:AURKA protein, human