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J Cell Biol ; 209(2): 221-34, 2015 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-25918224

RESUMO

Chromosome congression requires the stable attachment of microtubules to chromosomes mediated by the kinetochore, a large proteinaceous structure whose mechanism of assembly is unknown. In this paper, we present the finding that a protein called TRAMM (formerly known as TrappC12) plays a role in mitosis. Depletion of TRAMM resulted in noncongressed chromosomes and arrested cells in mitosis. Small amounts of TRAMM associated with chromosomes, and its depletion affected the localization of some kinetochore proteins, the strongest effect being seen for CENP-E. TRAMM interacts with CENP-E, and depletion of TRAMM prevented the recruitment of CENP-E to the kinetochore. TRAMM is phosphorylated early in mitosis and dephosphorylated at the onset of anaphase. Interestingly, this phosphorylation/dephosphorylation cycle correlates with its association/disassociation with CENP-E. Finally, we demonstrate that a phosphomimetic form of TRAMM recruited CENP-E to kinetochores more efficiently than did the nonphosphorylatable mutant. Our study identifies a moonlighting function for TRAMM during mitosis and adds a new component that regulates kinetochore stability and CENP-E recruitment.


Assuntos
Proteínas Cromossômicas não Histona/metabolismo , Segregação de Cromossomos , Cromossomos Humanos , Cinetocoros/química , Cinetocoros/metabolismo , Mitose/fisiologia , Proteínas de Transporte Vesicular/metabolismo , Western Blotting , Posicionamento Cromossômico , Células HeLa , Humanos , Microtúbulos/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fuso Acromático , Técnicas do Sistema de Duplo-Híbrido
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