title: Mutations in the paxillin-binding site of integrin-linked kinase (ILK) destabilize the pseudokinase domain and cause embryonic lethality in mice.
abstract: Integrin-linked kinase (ILK) localizes to focal adhesions (FAs) where it regulates cell spreading, migration, and growth factor receptor signaling. Previous reports showed that overexpressed ILK in which Val(386) and Thr(387) were substituted with glycine residues (ILK-VT/GG) could neither interact with paxillin nor localize to FA in cells expressing endogenous wild-type ILK, implying that paxillin binding to ILK is required for its localization to FAs. Here, we show that introducing this mutation into the germ line of mice (ILK-VT/GG) caused vasculogenesis defects, resulting in a general developmental delay and death at around embryonic day 12.5. Fibroblasts isolated from ILK-VT/GG mice contained mutant ILK in FAs, showed normal adhesion to and spreading on extracellular matrix substrates but displayed impaired migration. Biochemical analysis revealed that VT/GG substitutions decreased ILK protein stability leading to decreased ILK levels and reduced binding to paxillin and α-parvin. Because paxillin depletion did not affect ILK localization to FAs, the embryonic lethality and the in vitro migration defects are likely due to the reduced levels of ILK-VT/GG and diminished binding to parvins.
authors: Department of Molecular Medicine, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.
authors: Daniel
authors: D
authors: Moik
authors: Anika
authors: A
authors: Böttcher
authors: Tatiana
authors: T
authors: Makhina
authors: Carsten
authors: C
authors: Grashoff
authors: Nada
authors: N
authors: Bulus
authors: Roy
authors: R
authors: Zent
authors: Reinhard
authors: R
authors: Fässler
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D002465:Cell Movement; D004622:Embryo, Mammalian / Q000196:embryology*; D005434:Flow Cytometry; D022001:Focal Adhesions / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D005804:Genes, Lethal; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D009154:Mutation; D051419:Paxillin / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013997:Time Factors; D000099200:Scaffold Protein ILK
chemical_list: D008840:Microfilament Proteins; D051419:Paxillin; D017346:Protein Serine-Threonine Kinases; D000099200:Scaffold Protein ILK; C519774:Parva protein, mouse
keywords: Cell Adhesion; Cell Migration; Development; Embryonic Development; Integrin-linked Kinase; PINCH; Parvin; Paxillin; Protein Stability; Vasculogenesis
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