KANK2 links αVβ5 focal adhesions to microtubules and regulates sensitivity to microtubule poisons and cell migration

Paradžik, Mladen; Humphries, Jonathan D.; Stojanović, Nikolina; Nestić, Davor; Majhen, Dragomira; Dekanić, Ana; Samaržija, Ivana; Sedda, Delphine; Weber, Igor; Humphries, Martin J.; Ambriović-Ristov, Andreja (2020) KANK2 links αVβ5 focal adhesions to microtubules and regulates sensitivity to microtubule poisons and cell migration. Frontiers in Cell and Developmental Biology, 8 . ISSN 2296-634X

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Official URL: https://www.frontiersin.org/articles/10.3389/fcell...

Abstract

Integrins are heterodimeric glycoproteins that bind cells to extracellular matrix. Upon integrin clustering, multimolecular integrin adhesion complexes (IACs) are formed, creating links to the cell cytoskeleton. We have previously observed decreased cell migration and increased sensitivity to microtubule (MT) poisons, paclitaxel and vincristine, in the melanoma cell line MDA-MB-435S upon transfection with integrin alpha V specific siRNA, suggesting a link between adhesion and drug sensitivity. To elucidate the underlying mechanism, we determined alpha V-dependent changes in IAC composition. Using mass spectrometry (MS)-based proteomics, we analyzed the components of isolated IACs of MDA-MB-435S cells and two MDA-MB-435S-derived integrin alpha V specific shRNA-expressing cell clones with decreased expression of integrin alpha V. MS analysis showed that cells preferentially use integrin alpha V beta 5 for the formation of IACs. The differential analysis between MDA-MB-435S cells and clones with decreased expression of integrin alpha V identified key components of integrin alpha V beta 5 adhesion complexes as talins 1 and 2, alpha-actinins 1 and 4, filamins A and B, plectin and vinculin. The data also revealed decreased levels of several components of the cortical microtubule stabilization complex, which recruits MTs to adhesion sites (notably liprins alpha and beta, ELKS, LL5b, MACF1, KANK1, and KANK2), following alpha V knockdown. KANK2 knockdown in MDA-MB-435S cells mimicked the effect of integrin alpha V knockdown and resulted in increased sensitivity to MT poisons and decreased migration. Taken together, we conclude that KANK2 is a key molecule linking integrin alpha V beta 5 IACs to MTs, and enabling the actin-MT crosstalk that is important for both sensitivity to MT poisons and cell migration.

Item Type:

Article

Uncontrolled Keywords:

adhesome ; integrin alpha V beta 5 ; antitumor drug resistance, cell migration ; cortical microtubule stabilizing complex ; KANK2

Subjects:

NATURAL SCIENCES > Biology

Divisions:

Division of Molecular Biology

Projects:

Project title	Project leader	Project code	Project type
MOLEKULARNI MEHANIZMI POVEĆANJA OSJETLJIVOSTI NA PROTUTUMORSKE LIJEKOVE STANICA KARCINOMA DOJKE I MELANOMA ČOVJEKA UTIŠAVANJEM INTEGRINA-INSILCELL	Andreja Ambriović Ristov	IP-2013-11-2465	HRZZ
UNSPECIFIED	UNSPECIFIED	IP-2019-04-1577	UNSPECIFIED

Depositing User:

Andreja Ambriović Ristov

Date Deposited:

27 Mar 2020 11:03

URI:

http://fulir.irb.hr/id/eprint/5387

DOI:

10.3389/fcell.2020.00125