Integrin-linked kinase regulates interphase and mitotic microtubule dynamics.
Integrin-linked kinase (ILK) localizes to both focal adhesions and centrosomes in distinct multiprotein complexes. Its dual function as a kinase and scaffolding protein has been well characterized at focal adhesions, where it regulates integrin-mediated cell adhesion, spreading, migration and signaling. At the centrosomes, ILK regulates mitotic spindle organization and centrosome clustering. Our previous study showed various spindle defects after ILK knockdown or inhibition that suggested alteration in microtubule dynamics. Since ILK expression is frequently elevated in many cancer types, we investigated the effects of ILK overexpression on microtubule dynamics. We show here that overexpressing ILK in HeLa cells was associated with a shorter duration of mitosis and decreased sensitivity to paclitaxel, a chemotherapeutic agent that suppresses microtubule dynamics. Measurement of interphase microtubule dynamics revealed that ILK overexpression favored microtubule depolymerization, suggesting that microtubule destabilization could be the mechanism behind the decreased sensitivity to paclitaxel, which is known to stabilize microtubules. Conversely, the use of a small molecule inhibitor selective against ILK, QLT-0267, resulted in suppressed microtubule dynamics, demonstrating a new mechanism of action for this compound. We further show that treatment of HeLa cells with QLT-0267 resulted in higher inter-centromere tension in aligned chromosomes during mitosis, slower microtubule regrowth after cold depolymerization and the presence of a more stable population of spindle microtubules. These results demonstrate that ILK regulates microtubule dynamics in both interphase and mitotic cells.
Department of Integrative Oncology, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
Simin
S
Lim
Eiko
E
Kawamura
Andrew B
AB
Fielding
Mykola
M
Maydan
Shoukat
S
Dedhar
D000818:Animals; D001391:Azo Compounds / Q000494:pharmacology; D002503:Centromere / Q000187:drug effects / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007399:Interphase / Q000187:drug effects; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D008938:Mitosis / Q000187:drug effects; D017239:Paclitaxel / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology; D013997:Time Factors; D000099200:Scaffold Protein ILK
D001391:Azo Compounds; D017239:Paclitaxel; D017346:Protein Serine-Threonine Kinases; D011720:Pyrazoles; D000099200:Scaffold Protein ILK; C552362:QLT 0267