A small C-terminal sequence of Aurora B is responsible for localization and function.

Aurora B, a protein kinase required in mitosis, localizes to inner centromeres at metaphase and the spindle midzone in anaphase and is required for proper chromosome segregation and cytokinesis. Aurora A, a paralogue of Aurora B, localizes instead to centrosomes and spindle microtubules. Except for distinct N termini, Aurora B and Aurora A have highly similar sequences. We have combined small interfering RNA (siRNA) ablation of Aurora B with overexpression of truncation mutants to investigate the role of Aurora B sequence in its function. Reintroduction of Aurora B during siRNA treatment restored its localization and function. This permitted a restoration of function test to determine the sequence requirements for Aurora B targeting and function. Using this rescue protocol, neither N-terminal truncation of Aurora B unique sequence nor substitution with Aurora A N-terminal sequence affected Aurora B localization or function. Truncation of unique Aurora B C-terminal sequence from terminal residue 344 to residue 333 was without effect, but truncation to 326 abolished localization and function. Deletion of residues 326-333 completely abolished localization and blocked cells at prometaphase, establishing this sequence as critical to Aurora B function. Our findings thus establish a small sequence as essential for the distinct localization and function of Aurora B.

[1]  Ryoma Ohi,et al.  The Chromosomal Passenger Complex Is Required for Chromatin-Induced Microtubule Stabilization and Spindle Assembly , 2004, Cell.

[2]  William C Earnshaw,et al.  Borealin: a novel chromosomal passenger required for stability of the bipolar mitotic spindle. , 2004, The Journal of cell biology.

[3]  W. Earnshaw,et al.  Chromosomal passengers: Toward an integrated view of mitosis , 1991, Chromosoma.

[4]  J. Garin,et al.  The mammalian passenger protein TD-60 is an RCC1 family member with an essential role in prometaphase to metaphase progression. , 2003, Developmental cell.

[5]  William C Earnshaw,et al.  Survivin is required for stable checkpoint activation in taxol-treated HeLa cells , 2003, Journal of Cell Science.

[6]  J. Peters,et al.  The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint , 2003, The Journal of cell biology.

[7]  Stephen S. Taylor,et al.  Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores , 2003, The Journal of cell biology.

[8]  A. Venkitaraman,et al.  AURORA-A amplification overrides the mitotic spindle assembly checkpoint, inducing resistance to Taxol. , 2003, Cancer cell.

[9]  Erich A. Nigg,et al.  Centrosome aberrations: cause or consequence of cancer progression? , 2002, Nature Reviews Cancer.

[10]  Stéphanie Dutertre,et al.  On the role of aurora-A in centrosome function , 2002, Oncogene.

[11]  Jian Kuang,et al.  Aurora B kinase exists in a complex with survivin and INCENP and its kinase activity is stimulated by survivin binding and phosphorylation. , 2002, Molecular biology of the cell.

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

[13]  B. Stillman,et al.  Orc6 Involved in DNA Replication, Chromosome Segregation, and Cytokinesis , 2002, Science.

[14]  R. Bernards,et al.  A System for Stable Expression of Short Interfering RNAs in Mammalian Cells , 2002, Science.

[15]  P. Meraldi,et al.  Aurora‐A overexpression reveals tetraploidization as a major route to centrosome amplification in p53−/− cells , 2002, The EMBO journal.

[16]  K. Nasmyth,et al.  Evidence that the Ipl1-Sli15 (Aurora Kinase-INCENP) Complex Promotes Chromosome Bi-orientation by Altering Kinetochore-Spindle Pole Connections , 2002, Cell.

[17]  C. Allis,et al.  Mitotic Phosphorylation of Histone H3: Spatio-Temporal Regulation by Mammalian Aurora Kinases , 2002, Molecular and Cellular Biology.

[18]  Erich A Nigg,et al.  Aurora‐B phosphorylates Histone H3 at serine28 with regard to the mitotic chromosome condensation , 2002, Genes to cells : devoted to molecular & cellular mechanisms.

[19]  Anthony A. Hyman,et al.  Aurora-A kinase is required for centrosome maturation in Caenorhabditis elegans , 2001, The Journal of cell biology.

[20]  A. Murray,et al.  The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint. , 2001, Genes & development.

[21]  A. Murray,et al.  Lack of tension at kinetochores activates the spindle checkpoint in budding yeast , 2001, Current Biology.

[22]  C. Prigent,et al.  The non-catalytic domain of the Xenopus laevis auroraA kinase localises the protein to the centrosome. , 2001, Journal of cell science.

[23]  W. Earnshaw,et al.  INCENP is required for proper targeting of Survivin to the centromeres and the anaphase spindle during mitosis , 2001, Current Biology.

[24]  Stefan Dimitrov,et al.  Histone H3 phosphorylation and cell division , 2001, Oncogene.

[25]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[26]  Richard R. Adams,et al.  Essential Roles of Drosophila Inner Centromere Protein (Incenp) and Aurora B in Histone H3 Phosphorylation, Metaphase Chromosome Alignment, Kinetochore Disjunction, and Chromosome Segregation , 2001, The Journal of cell biology.

[27]  D. Glover,et al.  Drosophila Aurora B Kinase Is Required for Histone H3 Phosphorylation and Condensin Recruitment during Chromosome Condensation and to Organize the Central Spindle during Cytokinesis , 2001, The Journal of cell biology.

[28]  W. Earnshaw,et al.  Chromosomal passengers and the (aurora) ABCs of mitosis. , 2001, Trends in cell biology.

[29]  C. Mollinari,et al.  Human Survivin Is a Kinetochore-Associated Passenger Protein , 2000, The Journal of cell biology.

[30]  M. Mendoza,et al.  Incenp and an Aurora-like kinase form a complex essential for chromosome segregation and efficient completion of cytokinesis , 2000, Current Biology.

[31]  R. R. Adams,et al.  INCENP binds the Aurora-related kinase AIRK2 and is required to target it to chromosomes, the central spindle and cleavage furrow , 2000, Current Biology.

[32]  H. Horvitz,et al.  The survivin-like C. elegans BIR-1 protein acts with the Aurora-like kinase AIR-2 to affect chromosomes and the spindle midzone. , 2000, Molecular cell.

[33]  J. Bischoff,et al.  The Aurora/Ipl1p kinase family: regulators of chromosome segregation and cytokinesis. , 1999, Trends in cell biology.

[34]  C. Prigent,et al.  Aurora/Ipl1p-related kinases, a new oncogenic family of mitotic serine-threonine kinases. , 1999, Journal of cell science.

[35]  Clarence S. M. Chan,et al.  Sli15 Associates with the Ipl1 Protein Kinase to Promote Proper Chromosome Segregation in Saccharomyces cerevisiae , 1999, The Journal of cell biology.

[36]  A. Murray,et al.  The conserved protein kinase Ipl1 regulates microtubule binding to kinetochores in budding yeast. , 1999, Genes & development.

[37]  J. Schumacher,et al.  AIR-2: An Aurora/Ipl1-related Protein Kinase Associated with Chromosomes and Midbody Microtubules Is Required for Polar Body Extrusion and Cytokinesis in Caenorhabditis elegans Embryos , 1998, The Journal of cell biology.

[38]  R. Margolis,et al.  Colocalization of TD-60 and INCENP throughout G2 and mitosis: evidence for their possible interaction in signalling cytokinesis , 1998, Chromosoma.

[39]  Jian Kuang,et al.  Tumour amplified kinase STK15/BTAK induces centrosome amplification, aneuploidy and transformation , 1998, Nature Genetics.

[40]  Brian Schryver,et al.  A homologue of Drosophila aurora kinase is oncogenic and amplified in human colorectal cancers , 1998, The EMBO journal.

[41]  M. Tatsuka,et al.  AIM‐1: a mammalian midbody‐associated protein required for cytokinesis , 1998, The EMBO journal.

[42]  Clarence S. M. Chan,et al.  A Novel Mammalian, Mitotic Spindle–associated Kinase Is Related to Yeast and Fly Chromosome Segregation Regulators , 1997, The Journal of cell biology.

[43]  S. Kotani,et al.  Cell Cycle-dependent Expression and Spindle Pole Localization of a Novel Human Protein Kinase, Aik, Related to Aurora of Drosophila and Yeast Ipl1* , 1997, The Journal of Biological Chemistry.

[44]  D. Botstein,et al.  Isolation and characterization of chromosome-gain and increase-in-ploidy mutants in yeast. , 1993, Genetics.

[45]  R. Margolis,et al.  Telophase disc: a new mammalian mitotic organelle that bisects telophase cells with a possible function in cytokinesis. , 1991, Journal of cell science.

[46]  W. Earnshaw,et al.  The inner centromere protein (INCENP) antigens: movement from inner centromere to midbody during mitosis , 1987, The Journal of cell biology.