AIBp regulates mitotic entry and mitotic spindle assembly by controlling activation of both Aurora-A and Plk1

We previously reported that Aurora-A and the hNinein binding protein AIBp facilitate centrosomal structure maintenance and contribute to spindle formation. Here, we report that AIBp also interacts with Plk1, raising the possibility of functional similarity to Bora, which subsequently promotes Aurora-A–mediated Plk1 activation at Thr210 as well as Aurora-A activation at Thr288. In kinase assays, AIBp acts not only as a substrate but also as a positive regulator of both Aurora-A and Plk1. However, AIBp functions as a negative regulator to block phosphorylation of hNinein mediated by Aurora-A and Plk1. These findings suggest a novel AIBp-dependent regulatory machinery that controls mitotic entry. Additionally, knockdown of hNinein caused failure of AIBp to target the centrosome, whereas depletion of AIBp did not affect the localization of hNinein and microtubule nucleation. Notably, knockdown of AIBp in HeLa cells impaired both Aurora-A and Plk1 kinase, resulting in phenotypes with multiple spindle pole formation and chromosome misalignment. Our data show that depletion of AIBp results in the mis-localization of TACC3 and ch-TOG, but not CEP192 and CEP215, suggesting that loss of AIBp dominantly affects the Aurora-A substrate to cause mitotic aberrations. Collectively, our data demonstrate that AIBp contributes to mitotic entry and bipolar spindle assembly and may partially control localization, phosphorylation, and activation of both Aurora-A and Plk1 via hNinein during mitotic progression.

[1]  K. Tamai,et al.  The centrosomal protein Lats2 is a phosphorylation target of Aurora‐A kinase , 2004, Genes to cells : devoted to molecular & cellular mechanisms.

[2]  Viji M. Draviam,et al.  The ch-TOG/XMAP215 protein is essential for spindle pole organization in human somatic cells. , 2003, Genes & development.

[3]  J. B. Rattner,et al.  CEP110 and ninein are located in a specific domain of the centrosome associated with centrosome maturation. , 2002, Journal of cell science.

[4]  K. Rhee,et al.  CEP215 is involved in the dynein-dependent accumulation of pericentriolar matrix proteins for spindle pole formation , 2010, Cell cycle.

[5]  E. Nigg,et al.  Phosphorylation of Nlp by Plk1 negatively regulates its dynein-dynactin-dependent targeting to the centrosome , 2005, Journal of Cell Science.

[6]  David M. Glover,et al.  Polo-like kinases: conservation and divergence in their functions and regulation , 2009, Nature Reviews Molecular Cell Biology.

[7]  Chi-Ying F. Huang,et al.  Molecular characterization of human ninein protein: two distinct subdomains required for centrosomal targeting and regulating signals in cell cycle. , 2003, Biochemical and biophysical research communications.

[8]  L. Brunetto,et al.  Aurora-A and ch-TOG act in a common pathway in control of spindle pole integrity , 2008, Oncogene.

[9]  H. Saya,et al.  Aurora-A — A guardian of poles , 2005, Nature Reviews Cancer.

[10]  E. Nigg,et al.  Plk1 regulates mitotic Aurora A function through βTrCP-dependent degradation of hBora , 2008, Chromosoma.

[11]  David M. Glover,et al.  Centrosome biogenesis and function: centrosomics brings new understanding , 2007, Nature Reviews Molecular Cell Biology.

[12]  W. Earnshaw,et al.  The cellular geography of Aurora kinases , 2003, Nature Reviews Molecular Cell Biology.

[13]  Jun Zhou,et al.  Human Ninein is a Centrosomal Autoantigen Recognized by CREST Patient Sera and Plays a Regulatory Role in Microtubule Nucleation , 2004, Cell cycle.

[14]  Michael B. Yaffe,et al.  The Molecular Basis for Phosphodependent Substrate Targeting and Regulation of Plks by the Polo-Box Domain , 2003, Cell.

[15]  Paul Taylor,et al.  Inhibitors of Polo-like kinase reveal roles in spindle-pole maintenance , 2006, Nature chemical biology.

[16]  H. Maiato,et al.  Mitotic spindle multipolarity without centrosome amplification , 2014, Nature Cell Biology.

[17]  S. Doxsey.  Duplicating dangerously: linking centrosome duplication and aneuploidy. , 2002, Molecular cell.

[18]  R. Poon,et al.  Co-inhibition of polo-like kinase 1 and Aurora kinases promotes mitotic catastrophe , 2015, Oncotarget.

[19]  P. Meraldi,et al.  Polo-like kinase 1 regulates Nlp, a centrosome protein involved in microtubule nucleation. , 2003, Developmental cell.

[20]  Michael B Yaffe,et al.  Proteomic Screen Finds pSer/pThr-Binding Domain Localizing Plk1 to Mitotic Substrates , 2003, Science.

[21]  Roman Körner,et al.  Human TPX2 is required for targeting Aurora-A kinase to the spindle , 2002, The Journal of cell biology.

[22]  R. Medema,et al.  Bora and Aurora-A continue to activate Plk1 in mitosis , 2014, Journal of Cell Science.

[23]  S. Howng,et al.  Characterization and Functional Aspects of Human Ninein Isoforms that Regulated by Centrosomal Targeting Signals and Evidence for Docking Sites to Direct Gamma-Tubulin , 2006, Cell cycle.

[24]  Li Zhu.  Yeast GAL4 Two-Hybrid System , 1997 .

[25]  Chi-Ying F. Huang,et al.  Functional characterization of AIBp, a novel Aurora-A binding protein in centrosome structure and spindle formation. , 2010, International journal of oncology.

[26]  M. Bornens,et al.  The Respective Contributions of the Mother and Daughter Centrioles to Centrosome Activity and Behavior in Vertebrate Cells , 2000, The Journal of cell biology.

[27]  S. Howng,et al.  A novel ninein‐interaction protein, CGI‐99, blocks ninein phosphorylation by GSK3β and is highly expressed in brain tumors , 2004, FEBS letters.

[28]  J. Yates,et al.  Bora and the Kinase Aurora A Cooperatively Activate the Kinase Plk1 and Control Mitotic Entry , 2008, Science.

[29]  M. Bornens,et al.  Structure and duplication of the centrosome , 2007, Journal of Cell Science.

[30]  J. Caldwell,et al.  Human Cep192 Is Required for Mitotic Centrosome and Spindle Assembly , 2007, Current Biology.

[31]  Eiji Kohmura,et al.  Aurora-A Kinase Maintains the Fidelity of Early and Late Mitotic Events in HeLa Cells* , 2003, Journal of Biological Chemistry.

[32]  Péter Lénárt,et al.  Polo on the Rise-from Mitotic Entry to Cytokinesis with Plk1. , 2008, Developmental cell.

[33]  K. Ohashi,et al.  Furry Protein Promotes Aurora A-mediated Polo-like Kinase 1 Activation* , 2012, The Journal of Biological Chemistry.

[34]  J. Yates,et al.  Plk1- and β-TrCP–dependent degradation of Bora controls mitotic progression , 2008, The Journal of cell biology.

[35]  Erich A. Nigg,et al.  Polo-like kinases and the orchestration of cell division , 2004, Nature Reviews Molecular Cell Biology.

[36]  A. Hyman,et al.  Aurora A phosphorylation of TACC3/maskin is required for centrosome-dependent microtubule assembly in mitosis , 2005, The Journal of cell biology.

[37]  D. Berdnik,et al.  Mitotic activation of the kinase Aurora-A requires its binding partner Bora. , 2006, Developmental cell.

[38]  M. Bornens.  Centrosome composition and microtubule anchoring mechanisms. , 2002, Current opinion in cell biology.

[39]  Miho Ohsugi,et al.  The Plk1 target Kizuna stabilizes mitotic centrosomes to ensure spindle bipolarity , 2006, Nature Cell Biology.

[40]  Y. Terada,et al.  Phosphorylation of Threonine 210 and the Role of Serine 137 in the Regulation of Mammalian Polo-like Kinase* , 2002, The Journal of Biological Chemistry.

[41]  Chi-Ying F. Huang,et al.  hNinein is required for targeting spindle-associated protein Astrin to the centrosome during the S and G2 phases. , 2007, Experimental cell research.

[42]  K. Rhee,et al.  PLK1 phosphorylation of pericentrin initiates centrosome maturation at the onset of mitosis , 2011, The Journal of cell biology.

[43]  M. Ahmadian,et al.  The Centrosomal Adaptor TACC3 and the Microtubule Polymerase chTOG Interact via Defined C-terminal Subdomains in an Aurora-A Kinase-independent Manner* , 2013, The Journal of Biological Chemistry.

[44]  Stephen S. Taylor,et al.  Polo-like kinase-1 is activated by aurora A to promote checkpoint recovery , 2008, Nature.

[45]  I. Prior,et al.  Aurora A kinase activity is required for localization of TACC3/ch-TOG/clathrin inter-microtubule bridges. , 2011, Communicative & integrative biology.

[46]  Miho Ohsugi,et al.  LATS2–Ajuba complex regulates γ‐tubulin recruitment to centrosomes and spindle organization during mitosis , 2006, FEBS letters.

[47]  Erich A Nigg,et al.  Aurora kinases link chromosome segregation and cell division to cancer susceptibility. , 2004, Current opinion in genetics & development.

[48]  D. Cleveland,et al.  Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis , 2009, Nature Reviews Molecular Cell Biology.

[49]  H. Saya,et al.  NDEL1 Phosphorylation by Aurora-A Kinase Is Essential for Centrosomal Maturation, Separation, and TACC3 Recruitment , 2006, Molecular and Cellular Biology.

[50]  J. Walter,et al.  The Cep192-organized aurora A-Plk1 cascade is essential for centrosome cycle and bipolar spindle assembly. , 2014, Molecular cell.

[51]  Vaishali Shinde,et al.  Localization of human TACC3 to mitotic spindles is mediated by phosphorylation on Ser558 by Aurora A: a novel pharmacodynamic method for measuring Aurora A activity. , 2007, Cancer research.

[52]  J. Peters,et al.  The Small-Molecule Inhibitor BI 2536 Reveals Novel Insights into Mitotic Roles of Polo-like Kinase 1 , 2007, Current Biology.

[53]  Q. Jiang,et al.  The role of mitotic kinases in coupling the centrosome cycle with the assembly of the mitotic spindle , 2014, Journal of Cell Science.

[54]  Jens Gruber,et al.  The mitotic-spindle-associated protein astrin is essential for progression through mitosis , 2002, Journal of Cell Science.

[55]  S. Howng,et al.  Cloning and characterization of a novel human ninein protein that interacts with the glycogen synthase kinase 3beta. , 2000, Biochimica et biophysica acta.

[56]  S. Howng,et al.  Cloning and characterization of a novel human ninein protein that interacts with the glycogen synthase kinase 3β , 2000 .