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Phosphorylation of targeting protein for Xenopus kinesin-like protein 2 (TPX2) at threonine 72 in spindle assembly.
Shim SY, Perez de Castro I, Neumayer G, Wang J, Park SK, Sanada K, Nguyen MD.
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The human ortholog of the targeting protein for Xenopus kinesin-like protein 2 (TPX2) is a cytoskeletal protein that plays a major role in spindle assembly and is required for mitosis. During spindle morphogenesis, TPX2 cooperates with Aurora A kinase and Eg5 kinesin to regulate microtubule organization. TPX2 displays over 40 putative phosphorylation sites identified from various high-throughput proteomic screenings. In this study, we characterize the phosphorylation of threonine 72 (Thr(72)) in human TPX2, a residue highly conserved across species. We find that Cdk1/2 phosphorylate TPX2 in vitro and in vivo. Using homemade antibodies specific for TPX2 phosphorylated at Thr(72), we show that this phosphorylation is cell cycle-dependent and peaks at M phase. Endogenous TPX2 phosphorylated at Thr(72) does not associate with the mitotic spindle. Furthermore, ectopic GFP-TPX2 T72A preferentially concentrates on the spindle, whereas GFP-TPX2 WT distributes to both spindle and cytosol. The T72A mutant also increases the proportion of cells with multipolar spindles phenotype. This effect is associated with increased Aurora A activity and abnormally elongated spindles, indicative of higher Eg5 activity. In summary, we propose that phosphorylation of Thr(72) regulates TPX2 localization and impacts spindle assembly via Aurora A and Eg5.
Aguirre-Portolés,
Tpx2 controls spindle integrity, genome stability, and tumor development.
2012,
Pubmed Anand,
AURORA-A amplification overrides the mitotic spindle assembly checkpoint, inducing resistance to Taxol.
2003,
Pubmed Asteriti,
Aurora-A inactivation causes mitotic spindle pole fragmentation by unbalancing microtubule-generated forces.
2011,
Pubmed Asteriti,
The Aurora-A/TPX2 complex: a novel oncogenic holoenzyme?
2010,
Pubmed Bayliss,
Structural basis of Aurora-A activation by TPX2 at the mitotic spindle.
2003,
Pubmed
,
Xenbase Belenguer,
Mitosis-specific phosphorylation of nucleolin by p34cdc2 protein kinase.
1990,
Pubmed Bischoff,
A homologue of Drosophila aurora kinase is oncogenic and amplified in human colorectal cancers.
1998,
Pubmed Brunet,
Characterization of the TPX2 domains involved in microtubule nucleation and spindle assembly in Xenopus egg extracts.
2004,
Pubmed
,
Xenbase Carter,
A signature of chromosomal instability inferred from gene expression profiles predicts clinical outcome in multiple human cancers.
2006,
Pubmed Chen,
CDC25B mediates rapamycin-induced oncogenic responses in cancer cells.
2009,
Pubmed Chi,
Identification of CDK2 substrates in human cell lysates.
2008,
Pubmed Daub,
Kinase-selective enrichment enables quantitative phosphoproteomics of the kinome across the cell cycle.
2008,
Pubmed De Azevedo,
Inhibition of cyclin-dependent kinases by purine analogues: crystal structure of human cdk2 complexed with roscovitine.
1997,
Pubmed Dephoure,
A quantitative atlas of mitotic phosphorylation.
2008,
Pubmed Eckerdt,
Spindle pole regulation by a discrete Eg5-interacting domain in TPX2.
2008,
Pubmed
,
Xenbase Eckerdt,
Phosphorylation of TPX2 by Plx1 enhances activation of Aurora A.
2009,
Pubmed
,
Xenbase Eyers,
Regulation of Xenopus Aurora A activation by TPX2.
2004,
Pubmed
,
Xenbase Franz-Wachtel,
Global detection of protein kinase D-dependent phosphorylation events in nocodazole-treated human cells.
2012,
Pubmed Garrett,
hTPX2 is required for normal spindle morphology and centrosome integrity during vertebrate cell division.
2002,
Pubmed
,
Xenbase Giubettini,
Control of Aurora-A stability through interaction with TPX2.
2011,
Pubmed Grosstessner-Hain,
Quantitative phospho-proteomics to investigate the polo-like kinase 1-dependent phospho-proteome.
2011,
Pubmed Gruss,
Chromosome-induced microtubule assembly mediated by TPX2 is required for spindle formation in HeLa cells.
2002,
Pubmed
,
Xenbase Hanahan,
The hallmarks of cancer.
2000,
Pubmed Hanahan,
Hallmarks of cancer: the next generation.
2011,
Pubmed Havlícek,
Cytokinin-derived cyclin-dependent kinase inhibitors: synthesis and cdc2 inhibitory activity of olomoucine and related compounds.
1997,
Pubmed Heidebrecht,
p100: a novel proliferation-associated nuclear protein specifically restricted to cell cycle phases S, G2, and M.
1997,
Pubmed Hornbeck,
PhosphoSitePlus: a comprehensive resource for investigating the structure and function of experimentally determined post-translational modifications in man and mouse.
2012,
Pubmed Kapoor,
Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5.
2000,
Pubmed
,
Xenbase Kettenbach,
Quantitative phosphoproteomics identifies substrates and functional modules of Aurora and Polo-like kinase activities in mitotic cells.
2011,
Pubmed Kufer,
Human TPX2 is required for targeting Aurora-A kinase to the spindle.
2002,
Pubmed
,
Xenbase Ma,
Nucleolin functions in nucleolus formation and chromosome congression.
2007,
Pubmed Ma,
TPX2 regulates the localization and activity of Eg5 in the mammalian mitotic spindle.
2011,
Pubmed Malik,
Quantitative analysis of the human spindle phosphoproteome at distinct mitotic stages.
2009,
Pubmed Mayya,
Quantitative phosphoproteomic analysis of T cell receptor signaling reveals system-wide modulation of protein-protein interactions.
2009,
Pubmed Meijer,
Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5.
1997,
Pubmed
,
Xenbase Neumayer,
TPX2: of spindle assembly, DNA damage response, and cancer.
2014,
Pubmed Neumayer,
TPX2 impacts acetylation of histone H4 at lysine 16: implications for DNA damage response.
2014,
Pubmed Neumayer,
Targeting protein for xenopus kinesin-like protein 2 (TPX2) regulates γ-histone 2AX (γ-H2AX) levels upon ionizing radiation.
2012,
Pubmed
,
Xenbase Nguyen,
Deregulation of Cdk5 in a mouse model of ALS: toxicity alleviated by perikaryal neurofilament inclusions.
2001,
Pubmed Nousiainen,
Phosphoproteome analysis of the human mitotic spindle.
2006,
Pubmed Olsen,
Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis.
2010,
Pubmed Pérez de Castro,
Mitotic Stress and Chromosomal Instability in Cancer: The Case for TPX2.
2012,
Pubmed Perkins,
Probability-based protein identification by searching sequence databases using mass spectrometry data.
1999,
Pubmed Peter,
Identification of major nucleolar proteins as candidate mitotic substrates of cdc2 kinase.
1990,
Pubmed Petry,
Branching microtubule nucleation in Xenopus egg extracts mediated by augmin and TPX2.
2013,
Pubmed
,
Xenbase Schatz,
Importin alpha-regulated nucleation of microtubules by TPX2.
2003,
Pubmed
,
Xenbase Sharma,
Regulation of cyclin-dependent kinase 5 catalytic activity by phosphorylation.
1999,
Pubmed Shiromizu,
Identification of missing proteins in the neXtProt database and unregistered phosphopeptides in the PhosphoSitePlus database as part of the Chromosome-centric Human Proteome Project.
2013,
Pubmed Sillars-Hardebol,
TPX2 and AURKA promote 20q amplicon-driven colorectal adenoma to carcinoma progression.
2012,
Pubmed Skoufias,
Mitosis persists in the absence of Cdk1 activity when proteolysis or protein phosphatase activity is suppressed.
2007,
Pubmed Stewart,
Anaphase-promoting complex/cyclosome controls the stability of TPX2 during mitotic exit.
2005,
Pubmed Thompson,
Mechanisms of chromosomal instability.
2010,
Pubmed Trieselmann,
Ran modulates spindle assembly by regulating a subset of TPX2 and Kid activities including Aurora A activation.
2003,
Pubmed
,
Xenbase Tsai,
p35 is a neural-specific regulatory subunit of cyclin-dependent kinase 5.
1994,
Pubmed Weber,
Dual phosphoproteomics and chemical proteomics analysis of erlotinib and gefitinib interference in acute myeloid leukemia cells.
2012,
Pubmed Wittmann,
Localization of the kinesin-like protein Xklp2 to spindle poles requires a leucine zipper, a microtubule-associated protein, and dynein.
1998,
Pubmed
,
Xenbase Wittmann,
TPX2, A novel xenopus MAP involved in spindle pole organization.
2000,
Pubmed
,
Xenbase Zheng,
Region-specific expression of cyclin-dependent kinase 5 (cdk5) and its activators, p35 and p39, in the developing and adult rat central nervous system.
1998,
Pubmed Zhou,
Toward a comprehensive characterization of a human cancer cell phosphoproteome.
2013,
Pubmed
