Probing cell-division phenotype space and Polo-like kinase function using small molecules

Peters, Ulf; Cherian, Joseph; Kim, Jeffrey H.; Kwok, Benjamin H.; Kapoor, Tarun M.

doi:10.1038/nchembio826

Cited by 109 publications

(108 citation statements)

References 47 publications

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“…The compound ON01910 was reported to be a Plk1 inhibitor [37], but in subsequent studies no inhibition of purified Plk1 was observed up to 30 μM [38]. Furthermore, the cellular phenotypes induced by ON01910 were not characteristic for Plk1 inactivation [39,40]. More selective Plk1 inhibitors have recently become available, [Reviewed in [24]].…”

Section: Plk1 Inhibitors-more Monopolar Spindlesmentioning

confidence: 99%

Mitotic drivers—inhibitors of the Aurora B Kinase

Keen

Taylor

2009

Cancer Metastasis Rev

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Section: Plk1 Inhibitors-more Monopolar Spindlesmentioning

confidence: 99%

Mitotic drivers—inhibitors of the Aurora B Kinase

Keen

Taylor

2009

Cancer Metastasis Rev

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“…Cells were imaged live before and after addition of the Aurora B inhibitor hesperadin 17 (50 nM) or the Plk inhibitor BTO-1 (20 µM) 27 . The YFP/CFP emission ratio was calculated from images acquired at each timepoint.…”

Section: Cell Culture Transfection and Live Imagingmentioning

confidence: 99%

Midzone activation of aurora B in anaphase produces an intracellular phosphorylation gradient

Fuller

Lampson

Foley

et al. 2008

Nature

339

380

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Proper partitioning of the contents of a cell between two daughters requires integration of spatial and temporal cues. The anaphase array of microtubules that self-organize at the spindle midzone contributes to positioning the cell division plane midway between the segregating chromosomes 1 . How this signaling occurs over micron length-scales, from the midzone to the cell cortex, is not known. Here we examine the anaphase dynamics of protein phosphorylation by Aurora B kinase, a key mitotic regulator, using FRET-based sensors in living cells and immunofluorescence of native Aurora B substrates. Quantitative analysis of phosphorylation dynamics, using chromosome and centromere targeted sensors, reveals that changes are due primarily to position along the division axis rather than time. These dynamics result in the formation of a spatial phosphorylation gradient early in anaphase that is centered at the spindle midzone. This gradient depends on Aurora B targeting to a subpopulation of microtubules that activate it. Aurora kinase activity organizes the targeted microtubules to generate a structure based feedback loop. We propose that feedback between Aurora B kinase activation and midzone microtubules generates a gradient of posttranslational marks that provides spatial information for events in anaphase and cytokinesis. Keywordsanaphase; gradient; FRET; Aurora B; INCENP; histone H3; serine 10; mitotic spindle; Hesperadin; monopolar It is believed that self-organizing systems position the cleavage furrow, since experimental displacement of the anaphase spindle results in repositioning of the cleavage furrow within minutes 2 . While mitotic chromosomes are thought to generate gradients of Ran GTP that selforganize the prometaphase spindle 3 , this cannot be the only self-organizing signal in anaphase because cytokinesis can occur in the absence of chromatin 4,5 . Instead, the location of the cleavage furrow is coupled to the position of the spindle midzone where the Chromosome *Correspondence to: PTS (e-mail pts7h@virginia.edu) or MAL (e-mail lampson@sas.upenn.edu). $ Both authors contributed equally to this work Author Information: Development of the Aurora B and Plk phosphorylation sensors and FRET imaging and analysis were done in the Kapoor lab by MA Lampson, together with EA Foley. BG Fuller performed immunofluorescence experiments. SR Nitcher and P Tobelman performed the kinase assays and P-Lisa experiments, respectively. KV Le performed live imaging of Aurora B-GFP. BG Fuller and MA Lampson wrote the paper. To examine spatial patterns of Aurora B signaling during anaphase, we developed a strategy using FRET-based sensors that report quantitative changes in substrate phosphorylation in living cells. We adapted a sensor design 6 in which changes in intramolecular CFP-YFP FRET depend on changes in phosphorylation of an Aurora B substrate peptide, which is conserved among members of the kinesin-13 family 7 (Fig. 1a). To mimic localizations of endogenous Aurora B substrates 8 , sensors were targeted to ce...

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“…The congruent effects of BI 2536 and BTO-1 reduce the likelihood that observations could be attributed to an overlap in off-target activity and support the conclusion that both compounds disrupt cytokinesis via Plk1. Nevertheless, the currently available data do not exclude the possibility that BI 2536 and BTO-1 also inhibit kinases other than Plk1 at the concentrations used for in vivo studies (e.g., Plk2, Plk3 22,31 ). Such multi-target kinase inhibition may complicate the use of these compounds in some research applications, but could possibly enhance their utility as therapeutic agents.…”

Section: A Role For Plk1 In Cytokinesis Initiationmentioning

confidence: 99%