Uncovering new substrates for Aurora A kinase

Sardón, Teresa; Pache, Roland A.; Stein, Amelie; Molina, Henrik; Vernos, Isabelle; Aloy, Patrick

doi:10.1038/embor.2010.171

Cited by 61 publications

(54 citation statements)

References 32 publications

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“…P-Ser 10 -H3 initiates during G 2 in pericentric foci and spreads along the chromosome arms, thus serving as a hallmark of mitosis [18]. Previously derived consensus sequences for Aurora kinases suggested that, similar to P-Ser 10 -H3, Ser 83 -HP1γ might be a target of Aurora kinases [19]. Additional experiments demonstrated that the temporal pattern of P-Ser 83 -HP1γ was similar to both Aurora A and Aurora B (Figure 1C).…”

Section: Resultsmentioning

confidence: 99%

Functional impact of Aurora A-mediated phosphorylation of HP1γ at serine 83 during cell cycle progression

Grzenda

Leonard

Seo

et al. 2013

Epigenetics & Chromatin

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BackgroundPrevious elegant studies performed in the fission yeast Schizosaccharomyces pombe have identified a requirement for heterochromatin protein 1 (HP1) for spindle pole formation and appropriate cell division. In mammalian cells, HP1γ has been implicated in both somatic and germ cell proliferation. High levels of HP1γ protein associate with enhanced cell proliferation and oncogenesis, while its genetic inactivation results in meiotic and mitotic failure. However, the regulation of HP1γ by kinases, critical for supporting mitotic progression, remains to be fully characterized.ResultsWe report for the first time that during mitotic cell division, HP1γ colocalizes and is phosphorylated at serine 83 (Ser83) in G2/M phase by Aurora A. Since Aurora A regulates both cell proliferation and mitotic aberrations, we evaluated the role of HP1γ in the regulation of these phenomena using siRNA-mediated knockdown, as well as phosphomimetic and nonphosphorylatable site-directed mutants. We found that genetic downregulation of HP1γ, which decreases the levels of phosphorylation of HP1γ at Ser83 (P-Ser83-HP1γ), results in mitotic aberrations that can be rescued by reintroducing wild type HP1γ, but not the nonphosphorylatable S83A-HP1γ mutant. In addition, proliferation assays showed that the phosphomimetic S83D-HP1γ increases 5-ethynyl-2´-deoxyuridine (EdU) incorporation, whereas the nonphosphorylatable S83A-HP1γ mutant abrogates this effect. Genome-wide expression profiling revealed that the effects of these mutants on mitotic functions are congruently reflected in G2/M gene expression networks in a manner that mimics the on and off states for P-Ser83-HP1γ.ConclusionsThis is the first description of a mitotic Aurora A-HP1γ pathway, whose integrity is necessary for the execution of proper somatic cell division, providing insight into specific types of posttranslational modifications that associate to distinct functional outcomes of this important chromatin protein.

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Section: Resultsmentioning

confidence: 99%

Functional impact of Aurora A-mediated phosphorylation of HP1γ at serine 83 during cell cycle progression

Grzenda

Leonard

Seo

et al. 2013

Epigenetics & Chromatin

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“…These substrates comprise the consensus sequence for Aurora A ([R/K/N]-R-X-[S/T]-B where B is any hydrophobic residue with the exception of Pro) (Ferrari et al, 2005; Ohashi et al, 2006; Sardon et al, 2010). Peptides were ordered through Genscript.…”

Section: Methodsmentioning

confidence: 99%

Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2

Zorba

Buosi

Kutter

et al. 2014

eLife

114

150

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We elucidate the molecular mechanisms of two distinct activation strategies (autophosphorylation and TPX2-mediated activation) in human Aurora A kinase. Classic allosteric activation is in play where either activation loop phosphorylation or TPX2 binding to a conserved hydrophobic groove shifts the equilibrium far towards the active conformation. We resolve the controversy about the mechanism of autophosphorylation by demonstrating intermolecular autophosphorylation in a long-lived dimer by combining X-ray crystallography with functional assays. We then address the allosteric activation by TPX2 through activity assays and the crystal structure of a domain-swapped dimer of dephosphorylated Aurora A and TPX21−25. While autophosphorylation is the key regulatory mechanism in the centrosomes in the early stages of mitosis, allosteric activation by TPX2 of dephosphorylated Aurora A could be at play in the spindle microtubules. The mechanistic insights into autophosphorylation and allosteric activation by TPX2 binding proposed here, may have implications for understanding regulation of other protein kinases.DOI: http://dx.doi.org/10.7554/eLife.02667.001

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“…Aurora A and B are ubiquitously expressed and involved in cell proliferation in many cell types, while Aurora C is commonly found in testicular tissue and does not regulate mitosis in somatic or tumor cells (Marumoto et al 2005; Lok et al 2010; Shimomura et al 2010). The biology of Aurora kinases is still in its infancy, particularly with regard to their mechanism of action, as the identification of their substrates and correlating function is ongoing (Sardon et al 2010). Generally, Aurora kinases serve as key regulators of mitotic events such as centrosome maturation/separation, mitotic entry, microtubule spindle assembly, chromosome assembly/segregation, mitotic checkpoint activation, and cytokinesis (Lok et al 2010; Carvajal et al 2006; Marumoto et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Anticancer activity of the Aurora A kinase inhibitor MK-5108 in non-small-cell lung cancer (NSCLC) in vitro as monotherapy and in combination with chemotherapies

Chinn

Holland

Mack

2014

J Cancer Res Clin Oncol

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Purpose Aurora kinases are key regulators of mitotic events. Dysfunction of these kinases can cause polyploidy and chromosomal instability, a contributor to tumorigenesis. MK-5108 is a potent inhibitor of Aurora A kinase that has shown preclinical potent activity in malignancies of breast, cervical, colon, ovarian, and pancreatic origin. We sought to assess the preclinical efficacy of MK-5108 in a panel of non-small-cell lung cancer cell lines as a single agent and in combination with cisplatin and docetaxel. Methods Eleven lung cancer cell lines were studied. Growth inhibition by MK-5108 was assessed with short- and long-term MTT assays. Cell cycling was measured by flow cytometry. Immunoblotting was used to determine targeted activity of MK-5108 on Aurora A and downstream effects (TACC3 and Plk1). Efficacy of combination studies performed with cisplatin and docetaxel was evaluated by median effect analysis. Results All cell lines demonstrated sustained growth inhibition following MK-5108 at varying nanomolar concentrations. MK-5108 induced G2/M accumulation, polyploidy, and apoptosis (increased sub-G1/PARP cleavage). Levels of Aurora A, TACC3, and Plk1 diminished. Concurrent treatment of MK-5108 with cisplatin or docetaxel synergistically inhibited cell growth with the docetaxel combination performing better. When administered sequentially, treatment with docetaxel first followed by MK-5108 exhibited greater growth inhibition than the inverse; yet concurrent treatment remained superior. Conclusions MK-5108 has potent anti-proliferative activity in lung cancer cell lines alone and in combination with chemotherapies. Determining how best to integrate Aurora inhibitors into current lung cancer treatment regimens would be beneficial.

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Uncovering new substrates for Aurora A kinase

Cited by 61 publications

References 32 publications

Functional impact of Aurora A-mediated phosphorylation of HP1γ at serine 83 during cell cycle progression

Functional impact of Aurora A-mediated phosphorylation of HP1γ at serine 83 during cell cycle progression

Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2

Anticancer activity of the Aurora A kinase inhibitor MK-5108 in non-small-cell lung cancer (NSCLC) in vitro as monotherapy and in combination with chemotherapies

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