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Title: Non-catalytic signaling by pseudokinase ILK for regulating cell adhesion

Abstract

Dynamic communication between integrin-containing complexes (focal adhesions, FAs) and actin filaments is critical for regulating cell adhesion. Pseudokinase ILK plays a key role in this process but the underlying mechanism remains highly elusive. Here in this paper we show that by recruiting FA adaptors PINCH and Parvin into a heterotrimeric complex (IPP), ILK triggers F-actin filament bundling – a process known to generate force/mechanical signal to promote cytoskeleton reassembly and dynamic cell adhesion. Structural, biochemical, and functional analyses revealed that the F-actin bundling is orchestrated by two previously unrecognized WASP-Homology-2 actin binding motifs within IPP, one from PINCH and the other from Parvin. Strikingly, this process is also sensitized to Mg-ATP bound to the pseudoactive site of ILK and its dysregulation severely impairs stress fibers formation, cell spreading, and migration. These data identify a crucial mechanism for ILK, highlighting its uniqueness as a pseudokinase to transduce non-catalytic signal and regulate cell adhesion.

Authors:
 [1];  [1];  [2];  [1];  [3];  [1];  [2]
  1. Cleveland Clinic, Cleveland, OH (United States)
  2. Cleveland Clinic, Cleveland, OH (United States); Case Western Reserve Univ., Cleveland, OH (United States)
  3. Case Western Reserve Univ., Cleveland, OH (United States)
Publication Date:
Research Org.:
Cleveland Clinic, Cleveland, OH (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1483387
Grant/Contract Number:  
AC02-06CH11357; HL058758; 1S10RR026820-01
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Vaynberg, Julia, Fukuda, Koichi, Lu, Fan, Bialkowska, Katarzyna, Chen, Yinghua, Plow, Edward F., and Qin, Jun. Non-catalytic signaling by pseudokinase ILK for regulating cell adhesion. United States: N. p., 2018. Web. doi:10.1038/s41467-018-06906-7.
Vaynberg, Julia, Fukuda, Koichi, Lu, Fan, Bialkowska, Katarzyna, Chen, Yinghua, Plow, Edward F., & Qin, Jun. Non-catalytic signaling by pseudokinase ILK for regulating cell adhesion. United States. doi:10.1038/s41467-018-06906-7.
Vaynberg, Julia, Fukuda, Koichi, Lu, Fan, Bialkowska, Katarzyna, Chen, Yinghua, Plow, Edward F., and Qin, Jun. Fri . "Non-catalytic signaling by pseudokinase ILK for regulating cell adhesion". United States. doi:10.1038/s41467-018-06906-7. https://www.osti.gov/servlets/purl/1483387.
@article{osti_1483387,
title = {Non-catalytic signaling by pseudokinase ILK for regulating cell adhesion},
author = {Vaynberg, Julia and Fukuda, Koichi and Lu, Fan and Bialkowska, Katarzyna and Chen, Yinghua and Plow, Edward F. and Qin, Jun},
abstractNote = {Dynamic communication between integrin-containing complexes (focal adhesions, FAs) and actin filaments is critical for regulating cell adhesion. Pseudokinase ILK plays a key role in this process but the underlying mechanism remains highly elusive. Here in this paper we show that by recruiting FA adaptors PINCH and Parvin into a heterotrimeric complex (IPP), ILK triggers F-actin filament bundling – a process known to generate force/mechanical signal to promote cytoskeleton reassembly and dynamic cell adhesion. Structural, biochemical, and functional analyses revealed that the F-actin bundling is orchestrated by two previously unrecognized WASP-Homology-2 actin binding motifs within IPP, one from PINCH and the other from Parvin. Strikingly, this process is also sensitized to Mg-ATP bound to the pseudoactive site of ILK and its dysregulation severely impairs stress fibers formation, cell spreading, and migration. These data identify a crucial mechanism for ILK, highlighting its uniqueness as a pseudokinase to transduce non-catalytic signal and regulate cell adhesion.},
doi = {10.1038/s41467-018-06906-7},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {2018},
month = {10}
}

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Works referenced in this record:

Regulation of cell adhesion and anchorage-dependent growth by a new β1-integrin-linked protein kinase
journal, January 1996

  • Hannigan, Gregory E.; Leung-Hagesteijn, Chungyee; Fitz-Gibbon, Linda
  • Nature, Vol. 379, Issue 6560
  • DOI: 10.1038/379091a0

C. elegans PAT-4/ILK Functions as an Adaptor Protein within Integrin Adhesion Complexes
journal, May 2002


Structural Basis of Focal Adhesion Localization of LIM-only Adaptor PINCH by Integrin-linked Kinase
journal, December 2008

  • Yang, Yanwu; Wang, Xiaoxia; Hawkins, Cheryl A.
  • Journal of Biological Chemistry, Vol. 284, Issue 9
  • DOI: 10.1074/jbc.M805319200

The Structure of α-Parvin CH2-Paxillin LD1 Complex Reveals a Novel Modular Recognition for Focal Adhesion Assembly
journal, May 2008

  • Wang, Xiaoxia; Fukuda, Koichi; Byeon, In-Ja
  • Journal of Biological Chemistry, Vol. 283, Issue 30
  • DOI: 10.1074/jbc.M801270200

The WH2 Domain and Actin Nucleation: Necessary but Insufficient
journal, June 2016


Structure of the LKB1-STRAD-MO25 Complex Reveals an Allosteric Mechanism of Kinase Activation
journal, November 2009


Emerging roles of pseudokinases
journal, September 2006

  • Boudeau, Jérôme; Miranda-Saavedra, Diego; Barton, Geoffrey J.
  • Trends in Cell Biology, Vol. 16, Issue 9
  • DOI: 10.1016/j.tcb.2006.07.003

Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA
journal, January 2017


CHOPCHOP: a CRISPR/Cas9 and TALEN web tool for genome editing
journal, May 2014

  • Montague, Tessa G.; Cruz, José M.; Gagnon, James A.
  • Nucleic Acids Research, Vol. 42, Issue W1
  • DOI: 10.1093/nar/gku410

Characterisation of IRTKS, a novel IRSp53/MIM family actin regulator with distinct filament bundling properties
journal, April 2007

  • Millard, T. H.; Dawson, J.; Machesky, L. M.
  • Journal of Cell Science, Vol. 120, Issue 9
  • DOI: 10.1242/jcs.001776

HKL -3000: the integration of data reduction and structure solution – from diffraction images to an initial model in minutes
journal, July 2006

  • Minor, Wladek; Cymborowski, Marcin; Otwinowski, Zbyszek
  • Acta Crystallographica Section D Biological Crystallography, Vol. 62, Issue 8
  • DOI: 10.1107/S0907444906019949

Integrin signalling adaptors: not only figurants in the cancer story
journal, November 2010

  • Cabodi, Sara; del Pilar Camacho-Leal, Maria; Di Stefano, Paola
  • Nature Reviews Cancer, Vol. 10, Issue 12
  • DOI: 10.1038/nrc2967

The secret life of kinases: insights into non-catalytic signalling functions from pseudokinases
journal, June 2017

  • Jacobsen, Annette V.; Murphy, James M.
  • Biochemical Society Transactions, Vol. 45, Issue 3
  • DOI: 10.1042/BST20160331

Drosophila Integrin-Linked Kinase Is Required at Sites of Integrin Adhesion to Link the Cytoskeleton to the Plasma Membrane
journal, March 2001

  • Zervas, Christos G.; Gregory, Stephen L.; Brown, Nicholas H.
  • The Journal of Cell Biology, Vol. 152, Issue 5
  • DOI: 10.1083/jcb.152.5.1007

Integration of actin dynamics and cell adhesion by a three-dimensional, mechanosensitive molecular clutch
journal, June 2015

  • Case, Lindsay B.; Waterman, Clare M.
  • Nature Cell Biology, Vol. 17, Issue 8
  • DOI: 10.1038/ncb3191

Determining the structures of large proteins and protein complexes by NMR
journal, January 1998


Structure and function of focal adhesions
journal, February 2012


Functional plasticity of CH domains
journal, December 2001


Spire and Cordon-bleu: multifunctional regulators of actin dynamics
journal, October 2008

  • Renault, Louis; Bugyi, Beáta; Carlier, Marie-France
  • Trends in Cell Biology, Vol. 18, Issue 10
  • DOI: 10.1016/j.tcb.2008.07.008

The integrin adhesome: from genes and proteins to human disease
journal, March 2014

  • Winograd-Katz, Sabina E.; Fässler, Reinhard; Geiger, Benjamin
  • Nature Reviews Molecular Cell Biology, Vol. 15, Issue 4
  • DOI: 10.1038/nrm3769

ILK, PINCH and parvin: the tIPP of integrin signalling
journal, December 2005

  • Legate, Kyle R.; Montañez, Eloi; Kudlacek, Oliver
  • Nature Reviews Molecular Cell Biology, Vol. 7, Issue 1
  • DOI: 10.1038/nrm1789

How integrins control mammary epithelial differentiation: A possible role for the ILK-PINCH-Parvin complex
journal, May 2011


Integrin-linked kinase is an adaptor with essential functions during mouse development
journal, October 2009

  • Lange, Anika; Wickström, Sara A.; Jakobson, Madis
  • Nature, Vol. 461, Issue 7266
  • DOI: 10.1038/nature08468

TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts
journal, June 2009

  • Shen, Yang; Delaglio, Frank; Cornilescu, Gabriel
  • Journal of Biomolecular NMR, Vol. 44, Issue 4
  • DOI: 10.1007/s10858-009-9333-z

PROCHECK: a program to check the stereochemical quality of protein structures
journal, April 1993

  • Laskowski, R. A.; MacArthur, M. W.; Moss, D. S.
  • Journal of Applied Crystallography, Vol. 26, Issue 2
  • DOI: 10.1107/S0021889892009944

Actin-bound structures of Wiskott-Aldrich syndrome protein (WASP)-homology domain 2 and the implications for filament assembly
journal, November 2005

  • Chereau, D.; Kerff, F.; Graceffa, P.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 46
  • DOI: 10.1073/pnas.0507021102

ILK: a pseudokinase in the center stage of cell-matrix adhesion and signaling
journal, October 2012


The structural basis of integrin-linked kinase-PINCH interactions
journal, December 2008

  • Chiswell, B. P.; Zhang, R.; Murphy, J. W.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 52
  • DOI: 10.1073/pnas.0811415106

Purification and SAXS Analysis of the Integrin Linked Kinase, PINCH, Parvin (IPP) Heterotrimeric Complex
journal, January 2013


Expression of a Nonpolymerizable Actin Mutant in Sf9 Cells
journal, September 2004

  • Joel, Peteranne B.; Fagnant, Patricia M.; Trybus, Kathleen M.
  • Biochemistry, Vol. 43, Issue 36
  • DOI: 10.1021/bi048899a

Solution Structure of the Focal Adhesion Adaptor PINCH LIM1 Domain and Characterization of Its Interaction with the Integrin-linked Kinase Ankyrin Repeat Domain
journal, November 2000

  • Velyvis, Algirdas; Yang, Yanwu; Wu, Chuanyue
  • Journal of Biological Chemistry, Vol. 276, Issue 7
  • DOI: 10.1074/jbc.M007632200

NMRPipe: A multidimensional spectral processing system based on UNIX pipes
journal, November 1995

  • Delaglio, Frank; Grzesiek, Stephan; Vuister, GeertenW.
  • Journal of Biomolecular NMR, Vol. 6, Issue 3
  • DOI: 10.1007/BF00197809

A Modular Polycistronic Expression System for Overexpressing Protein Complexes in Escherichia coli
journal, February 2001


The Xplor-NIH NMR molecular structure determination package
journal, January 2003


Integrin-linked kinase: a cancer therapeutic target unique among its ILK
journal, January 2005

  • Hannigan, Gregory; Troussard, Armelle A.; Dedhar, Shoukat
  • Nature Reviews Cancer, Vol. 5, Issue 1
  • DOI: 10.1038/nrc1524

Biochemical, Proteomic, Structural, and Thermodynamic Characterizations of Integrin-linked Kinase (ILK): CROSS-VALIDATION OF THE PSEUDOKINASE
journal, April 2011

  • Fukuda, Koichi; Knight, James D. R.; Piszczek, Grzegorz
  • Journal of Biological Chemistry, Vol. 286, Issue 24
  • DOI: 10.1074/jbc.M111.240093

The Pseudoactive Site of ILK Is Essential for Its Binding to α-Parvin and Localization to Focal Adhesions
journal, December 2009


Structure of an Ultraweak Protein-Protein Complex and Its Crucial Role in Regulation of Cell Morphology and Motility
journal, February 2005


A common sense approach to peak picking in two-, three-, and four-dimensional spectra using automatic computer analysis of contour diagrams
journal, December 2011

  • Garrett, Daniel S.; Powers, Robert; Gronenborn, Angela M.
  • Journal of Magnetic Resonance, Vol. 213, Issue 2
  • DOI: 10.1016/j.jmr.2011.09.007

MOLREP an Automated Program for Molecular Replacement
journal, December 1997


Zebrafish integrin-linked kinase is required in skeletal muscles for strengthening the integrin–ECM adhesion complex
journal, June 2008


Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to make actin bundles in cells
journal, March 2006

  • Loomis, Patricia A.; Kelly, Alexander E.; Zheng, Lili
  • Journal of Cell Science, Vol. 119, Issue 8
  • DOI: 10.1242/jcs.02869

Structure of a Bud6/Actin Complex Reveals a Novel WH2-like Actin Monomer Recruitment Motif
journal, August 2015


The ILK/PINCH/parvin complex: the kinase is dead, long live the pseudokinase!
journal, December 2009

  • Wickström, Sara A.; Lange, Anika; Montanez, Eloi
  • The EMBO Journal, Vol. 29, Issue 2
  • DOI: 10.1038/emboj.2009.376

Mechanotransduction and extracellular matrix homeostasis
journal, October 2014

  • Humphrey, Jay D.; Dufresne, Eric R.; Schwartz, Martin A.
  • Nature Reviews Molecular Cell Biology, Vol. 15, Issue 12
  • DOI: 10.1038/nrm3896

The regulation of integrin-linked kinase in human platelets: evidence for involvement in the regulation of integrin alpha2beta1
journal, August 2004