Fuller, B. G. et al. Midzone activation of Aurora B in anaphase produces an intracellular phosphorylation gradient. Nature 453, 1132-1136

Department of Biochemistry, University of Virginia School of Medicine, Charlottesville, Virginia 22908, USA.
Nature (Impact Factor: 41.46). 07/2008; 453(7198):1132-6. DOI: 10.1038/nature06923
Source: PubMed


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. How this signalling occurs over length scales of micrometres, 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 fluorescence resonance energy transfer (FRET)-based sensors in living HeLa 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 centred 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 post-translational marks that provides spatial information for events in anaphase and cytokinesis.

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Available from: Kim Le, Oct 05, 2015
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    • "Finally, KIF20A is responsible for recruiting the CPC to the spindle midzone, thereby creating a pool of active Aurora B, which is essential for controlling the activity of KIF4A and KIF23 (Gruneberg et al. 2004; Fuller et al. 2008). In addition to these factors, other MAPs are likely to be involved in central spindle assembly and maintenance (Table 1), including the kinesin KIF14, which interacts with PRC1 (Gruneberg et al. 2006; Bassi et al. 2013), KIF18 (Gatt et al. 2005), CLIP-associating protein 1 (CLASP) (Inoue et al. 2004), and abnormal spindlelike microcephaly-associated protein (ASPM), which is the only MAP that localizes to the minus ends of central spindle microtubules (Wakefield et al. 2001; Riparbelli et al. 2002). "
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    ABSTRACT: Cell division ends with the physical separation of the two daughter cells, a process known as cytokinesis. This final event ensures that nuclear and cytoplasmic contents are accurately partitioned between the two nascent cells. Cytokinesis is one of the most dramatic changes in cell shape and requires an extensive reorganization of the cell's cytoskeleton. Here, we describe the cytoskeletal structures, factors, and signaling pathways that orchestrate this robust and yet highly dynamic process in animal cells. Finally, we discuss possible future directions in this growing area of cell division research and its implications in human diseases, including cancer. Copyright © 2015 Cold Spring Harbor Laboratory Press; all rights reserved.
    Cold Spring Harbor perspectives in biology 02/2015; 7(4). DOI:10.1101/cshperspect.a015834 · 8.68 Impact Factor
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    • "Protein kinase Donor/acceptor fluorescent Phosphorylation site Phosphorylation binding domain Refs. proteins Cyclin B1-Cdk1 mcerulean/YPet Ser 126 of HsCyclin B1 Polo-box domain of HsPlk1 [19] Aurora-B CyPet/YPet or mTFP1/YPet Thr 67 of HsKif2 FHA2 of ScRad53 [56] [61] Plk1 CyPet/YPet Ser 426 of HsMyt1 FHA2 of ScRad53 [56] Plk1 CyPet/YPet or mTFP1/YPet Ser 17 of c-jun FHA2 of ScRad53 [59] Protein Name of the biosensor Donor/acceptor fluorescent Other domains Refs. proteins ran Rango Cerulean/EYFP Importin-β binding domain [40] of Hs snurportin1 Stathmin/Op18 COPY ECFP/citrine Full length Stathmin/Op18 [54] RhoA Raichu-RhoA SEYFP/SECFP Truncated RhoA and RhoA [73] "
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    ABSTRACT: Mitosis has been studied since the early 1880s as a key event of the cell division cycle where remarkable changes in cellular architecture take place and ultimately lead to an equal segregation of duplicated chromosomes into two daughter cells. A detailed description of the complex and highly ordered cellular events taking place is now available. Many regulators involved in key steps including entry into mitosis, nuclear envelope breakdown, microtubule (MT) spindle formation, and chromosome attachment, as well as mitotic exit and cytokinesis, have also been identified. However, understanding the precise spatio-temporal contribution of each regulator in the cell reorganization process has been technically challenging. This review will focus on a number of recent advances in our understanding of the spatial distribution of protein activities and the temporal regulation of their activation and inactivation during entry and progression through mitosis by the use of intramolecular Förster resonance energy transfer (FRET)-based biosensors.
    Biotechnology Journal 02/2014; 9(2). DOI:10.1002/biot.201300194 · 3.49 Impact Factor
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    • "Aurora B is found at the spindle midzone and at the equatorial cortex during the meta-to-anaphase transition [4]. At the spindle midzone, Aurora B is thought to generate an anaphase phosphorylation gradient toward the cell cortex, which provides spatial information to position the cleavage furrow [5]. In contrast, the importance of cortically localized Aurora B for cytokinesis has remained elusive. "
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    ABSTRACT: Although Aurora B is important in cleavage furrow ingression and completion during cytokinesis, the mechanism by which kinase activity is targeted to the cleavage furrow and the molecule(s) responsible for this process have remained elusive. Here, we demonstrate that an essential mitotic kinesin MKlp2 requires myosin-II for its localization to the equatorial cortex, and this event is required to recruit Aurora B to the equatorial cortex in mammalian cells. This recruitment event is also required to promote the highly focused accumulation of active RhoA at the equatorial cortex and stable ingression of the cleavage furrow in bipolar cytokinesis. Specifically, in drug-induced monopolar cytokinesis, targeting Aurora B to the cell cortex by MKlp2 is essential for cell polarization and furrow formation. Once the furrow has formed, MKlp2 further recruits Aurora B to the growing furrow. This process together with continuous Aurora B kinase activity at the growing furrow is essential for stable furrow propagation and completion. In contrast, a MKlp2 mutant defective in binding myosin-II does not recruit Aurora B to the cell cortex and does not promote furrow formation during monopolar cytokinesis. This mutant is also defective in maintaining the ingressing furrow during bipolar cytokinesis. Together, these findings reveal that targeting Aurora B to the cell cortex (or the equatorial cortex) by MKlp2 is essential for the maintenance of the ingressing furrow for successful cytokinesis.
    PLoS ONE 06/2013; 8(6):e64826. DOI:10.1371/journal.pone.0064826 · 3.23 Impact Factor
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