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MBC in Press, published online ahead of print October 27, 2004
Mol. Biol. Cell 10.1091/mbc.E04-06-0447

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Submitted on June 4, 2004
Revised on September 22, 2004
Accepted on October 13, 2004

A Small C-Terminal Sequence of Aurora B Is Responsible for Localization and Function

Laetitia Scrittori,*{dagger} Dimitrios Skoufias,{dagger}{ddagger} Fabienne Hans,*{dagger} Véronique Gerson,* Paolo Sassone-Corsi,{sect} Stefan Dimitrov,*|| and Robert Margolis{ddagger}||

*Institut Albert Bonniot, Institut National de la Santé et de la Recherche Médicale, 38706 La Tronche cedex, France; {ddagger}Institut de Biologie Structurale Jean Pierre Ebel (Commissariat à l’Energie Atomique/Centre National de la Recherche Scientifique), 38027 Grenoble cedex 1, France; {sect}Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique - Institut National de la Santé et de la Recherche Médicale -Université Louis Pasteur, 67404 Illkirch-Strasbourg, France

Monitoring Editor: J. Richard McIntosh

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 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.

{dagger}These authors contributed equally to this work.

||Corresponding authors. E-mail: stefan.dimitrov{at}ujf-grenoble.fr E-mail: Margolis{at}ibs.fr






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