Abstract
Integrin-linked kinase (ILK) is a serine-threonine kinase and scaffold protein with well defined roles in focal adhesions in integrin-mediated cell adhesion, spreading, migration, and signaling. Using mass spectrometry-based proteomic approaches, we identify centrosomal and mitotic spindle proteins as interactors of ILK. alpha- and beta-tubulin, ch-TOG (XMAP215), and RUVBL1 associate with ILK and colocalize with it to mitotic centrosomes. Inhibition of ILK activity or expression induces profound apoptosis-independent defects in the organization of the mitotic spindle and DNA segregation. ILK fails to localize to the centrosomes of abnormal spindles in RUVBL1-depleted cells. Additionally, depletion of ILK expression or inhibition of its activity inhibits Aurora A-TACC3/ch-TOG interactions, which are essential for spindle pole organization and mitosis. These data demonstrate a critical and unexpected function for ILK in the organization of centrosomal protein complexes during mitotic spindle assembly and DNA segregation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ATPases Associated with Diverse Cellular Activities
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Aurora Kinases
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Carrier Proteins / chemistry
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Centrosome / enzymology*
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Centrosome / ultrastructure
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Chromosome Segregation / genetics*
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DNA Helicases / chemistry
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DNA Helicases / genetics
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DNA Helicases / metabolism
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Down-Regulation / genetics
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HeLa Cells
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Humans
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Mass Spectrometry
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Microtubule-Associated Proteins / chemistry
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Microtubule-Associated Proteins / metabolism
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Mitosis / genetics*
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism*
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Protein Serine-Threonine Kinases / chemistry
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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Protein Structure, Tertiary / genetics
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Spindle Apparatus / enzymology*
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Spindle Apparatus / genetics
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Spindle Apparatus / ultrastructure
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Tubulin / chemistry
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Tubulin / metabolism
Substances
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CKAP5 protein, human
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Carrier Proteins
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Microtubule-Associated Proteins
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Nuclear Proteins
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Tubulin
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integrin-linked kinase
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Aurora Kinases
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Protein Serine-Threonine Kinases
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ATPases Associated with Diverse Cellular Activities
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DNA Helicases
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RUVBL1 protein, human