The development of multicellular organisms requires integrin-mediated interactions between cells and their extracellular environment. Integrin binding to extracellular matrix catalyses assembly of multiprotein complexes, which transduce mechanical and chemical signals that regulate many aspects of cell physiology1,2. Integrin-linked kinase (Ilk) is a multifunctional protein that binds β-integrin cytoplasmic domains and regulates actin dynamics by recruiting actin binding regulatory proteins such as α- and β-parvin3. Ilk has also been shown to possess serine/threonine kinase activity4 and to phosphorylate signalling proteins such as Akt1 and glycogen synthase kinase 3β (Gsk3β) in mammalian cells5; however, these functions have been shown by genetic studies6,7 not to occur in flies and worms. Here we show that mice carrying point mutations in the proposed autophosphorylation site of the putative kinase domain and in the pleckstrin homology domain are normal. In contrast, mice with point mutations in the conserved lysine residue of the potential ATP-binding site of the kinase domain, which mediates Ilk binding to α-parvin, die owing to renal agenesis. Similar renal defects occur in α-parvin-null mice. Thus, we provide genetic evidence that the kinase activity of Ilk is dispensable for mammalian development; however, an interaction between Ilk and α-parvin is critical for kidney development.
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We thank M. Bösl and M. Moser for help with the generation of the mutant mice, E. Montanez for help with the analysis of the α-parvin-null mice, M. Grzejszczyk and S. Bach for technical assistance, O. Kudlacek for help with initial vector construction, M. Aumailley (Univ. Cologne) for laminin322, A. A. Noegel (Univ. Cologne) for α-parvin cDNA, A. Miettinen (Univ. Helsinki) for the brush border antibody, and R. Böttcher for careful reading of the manuscript. This work was supported by the Sigrid Juselius Foundation (to S.A.W. and M.J.), the Academy of Finland (to S.A.W., M.J. and K.S.), the Finnish Cultural Foundation (to S.A.W.), NIDDK DK065123, DK075594, DK65123, an AHA established investigator award and a Merit award from the Department of Veterans Affairs (to R.Z.), the Austrian Science Funds (grant SFB021) and the Max Planck Society (to R.F.).
Author Contributions R.F. initiated and supervised the project. A.L., S.A.W., M.J., R.Z., K.S. and R.F. conceived ideas, designed experiments and analysed data. A.L., S.A.W., M.J. and R.Z. performed experiments. A.L., S.A.W., R.Z. and R.F. wrote the manuscript. All authors edited and reviewed the manuscript.
This file contains Supplementary Figures S1-S13 with Legends and Legends for Supplementary Movies 1-2. (PDF 14087 kb)
This movie shows the analysis of focal adhesion and actin dynamics in wild type Ilk-Flag cells using TIRF (see file s1 for full Legend). (MOV 4703 kb)
This movie shows the analysis of focal adhesion and actin dynamics in Ilk(K220A)-Flag cells using TIRF (see file S1 for full Legend). (MOV 5616 kb)
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Lange, A., Wickström, S., Jakobson, M. et al. Integrin-linked kinase is an adaptor with essential functions during mouse development. Nature 461, 1002–1006 (2009). https://doi.org/10.1038/nature08468
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