The interaction between uPAR and vitronectin triggers ligand ‐ independent adhesion signalling by integrins by Gian Maria Sarra Ferraris, Carsten Schulte,

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The interaction between uPAR and vitronectin triggers ligand ‐ independent adhesion signalling by integrins by Gian Maria Sarra Ferraris, Carsten Schulte, Valentina Buttiglione, Valentina De Lorenzi, Andrea Piontini, Massimiliano Galluzzi, Alessandro Podestà, Chris D Madsen, and Nicolai Sidenius EMBO J. Volume ():e August 28, 2014 ©2014 by European Molecular Biology Organization

uPAR ‐ mediated VN adhesion triggers signalling and cell spreading through αvβ5 and β1 integrins AVN adhesion of cells expressing uPAR variants in the presence or absence of uPA. Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

The RGD motif VN is required for integrin ‐ mediated cell adhesion but dispensable for uPAR ‐ induced β1 integrin signalling and cell spreading AEffect of VN variants on uPAR and integrin ‐ dependent cell adhesion. 293 uPART54A cells were seeded, in the presenc... Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

Localisation of β1 integrin and focal adhesion components during uPAR ‐ mediated cell spreading A–C293 uPART54A were seeded on FN or VNRAD in the presence of uPA for 30 min and stained with the indicated antibodies. Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

Structure–function analysis of the β1 integrin in uPAR/VN ‐ induced cell spreading and signalling A, BEffect of β1 integrin mutants on canonical integrin ‐ mediated FN adhesion and signalling. 293 cells co ‐ expressing uPART54A and the indicated β1 variants wer... Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

Ligand ‐ independent integrin signalling by β3 integrin Adhesive properties of 293 uPART54A cells expressing β3 integrin mutants. Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

Structural requirements to the anchoring receptor, its ligand and type of membrane anchorage Matrix ‐ binding activity is required for the induction of intracellular signalling and spreading. 293 cells expressing an engineered cell surface VN receptor (PAI ‐ 1... Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

uPAR/VN signalling requires cell adhesion, ECM rigidity and membrane tension uPAR ‐ induced signalling requires cell attachment. Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization

Comparison between canonical and ligand ‐ independent integrin signallingIn canonical signalling, integrins engage the specific ligands in the ECM and signal by recruiting proteins on the cytoplasmic tails. Gian Maria Sarra Ferraris et al. EMBO J. doi: /embj ©2014 by European Molecular Biology Organization