Aurora-A kinase-inactive mutants disrupt the interaction with Ajuba and cause defects in mitotic spindle formation and G2/M phase arrest in HeLa cells
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Aurora-A is a centrosome-localized serine/threonine kinasethat is overexpressed in multiple human cancers. We previouslyreported an intramolecular inhibitory regulation ofAurora-A between its N-terminal regulatory domain (Nt, aminoacids [aa] 1-128) and the C-terminal catalytic domain (Cd, aa129-403). Here, we demonstrate that although both Aurora-Amutants (AurA-K250G and AurA-D294G/Y295G) lackedinteractions between the Nt and Cd, they also failed to interactwith Ajuba, an essential activator of Aurora-A, leading to lossof kinase activity. Additionally, overexpression of either of themutants resulted in centrosome amplification and mitoticspindle formation defects. Both mutants were also able tocause G2/M arrest and apoptosis. These results indicate thatboth K250 and D294/Y295 are critical for direct interactionbetween Aurora-A and Ajuba and the function of the Aurora-Acomplex in cell cycle progression.
Abstract
Aurora-A is a centrosome-localized serine/threonine kinasethat is overexpressed in multiple human cancers. We previouslyreported an intramolecular inhibitory regulation ofAurora-A between its N-terminal regulatory domain (Nt, aminoacids [aa] 1-128) and the C-terminal catalytic domain (Cd, aa129-403). Here, we demonstrate that although both Aurora-Amutants (AurA-K250G and AurA-D294G/Y295G) lackedinteractions between the Nt and Cd, they also failed to interactwith Ajuba, an essential activator of Aurora-A, leading to lossof kinase activity. Additionally, overexpression of either of themutants resulted in centrosome amplification and mitoticspindle formation defects. Both mutants were also able tocause G2/M arrest and apoptosis. These results indicate thatboth K250 and D294/Y295 are critical for direct interactionbetween Aurora-A and Ajuba and the function of the Aurora-Acomplex in cell cycle progression.
참고문헌 (18)
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