title: Biochemical and functional characterization of the interaction between liprin-α1 and GIT1: implications for the regulation of cell motility.
abstract: We have previously identified the scaffold protein liprin-α1 as an important regulator of integrin-mediated cell motility and tumor cell invasion. Liprin-α1 may interact with different proteins, and the functional significance of these interactions in the regulation of cell motility is poorly known. Here we have addressed the involvement of the liprin-α1 partner GIT1 in liprin-α1-mediated effects on cell spreading and migration. GIT1 depletion inhibited spreading by affecting the lamellipodia, and prevented liprin-α1-enhanced spreading. Conversely inhibition of the formation of the liprin-α1-GIT complex by expression of liprin-ΔCC3 could still enhance spreading, although to a lesser extent compared to full length liprin-α1. No cumulative effects were observed after depletion of both liprin-α1 and GIT1, suggesting that the two proteins belong to the same signaling network in the regulation of cell spreading. Our data suggest that liprin-α1 may compete with paxillin for binding to GIT1, while binding of βPIX to GIT1 was unaffected by the presence of liprin-α1. Interestingly, GIT and liprin-α1 reciprocally regulated their subcellular localization, since liprin-α1 overexpression, but not the GIT binding-defective liprin-ΔCC3 mutant, affected the localization of endogenous GIT at peripheral and mature central focal adhesions, while the expression of a truncated, active form of GIT1 enhanced the localization of endogenous liprin-α1 at the edge of spreading cells. Moreover, GIT1 was required for liprin-α1-enhanced haptotatic migration, although the direct interaction between liprin-α1 and GIT1 was not needed. Our findings show that the functional interaction between liprin-α1 and GIT1 cooperate in the regulation of integrin-dependent cell spreading and motility on extracellular matrix. These findings and the possible competition of liprin-α1 with paxillin for binding to GIT1 suggest that alternative binding of GIT1 to either liprin-α1 or paxillin plays distinct roles in different phases of the protrusive activity in the cell.
authors.affiliation: Division of Neuroscience, San Raffaele Scientific Institute and San Raffaele University, Milano, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Asperti
authors.forename: Veronica
authors.initials: V
authors.lastname: Astro
authors.forename: Emanuela
authors.initials: E
authors.lastname: Pettinato
authors.forename: Simona
authors.initials: S
authors.lastname: Paris
authors.forename: Angela
authors.initials: A
authors.lastname: Bachi
authors.forename: Ivan
authors.initials: I
authors.lastname: de Curtis
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002465:Cell Movement; D048430:Cell Shape; D002522:Chlorocebus aethiops; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051419:Paxillin / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D064067:Rho Guanine Nucleotide Exchange Factors; D013347:Subcellular Fractions
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D020662:Guanine Nucleotide Exchange Factors; D051419:Paxillin; D064067:Rho Guanine Nucleotide Exchange Factors