Abstract:
:The molecular clutch (MC) model proposes that actomyosin-driven force transmission permits integrin-dependent cell migration. To investigate the MC, we introduced diverse talin (TLN) and integrin variants into Flp-In™ T-Rex™ HEK293 cells stably expressing uPAR. Vitronectin variants served as substrate providing uPAR-mediated cell adhesion and optionally integrin binding. This particular system allowed us to selectively analyse key MC proteins and interactions, effectively from the extracellular matrix substrate to intracellular f-actin, and to therewith study mechanobiological aspects of MC engagement also uncoupled from integrin/ligand binding. With this experimental approach, we found that for the initial PIP2-dependent membrane/TLN/f-actin linkage and persistent lamellipodia formation the C-terminal TLN actin binding site (ABS) is dispensable. The establishment of an adequate MC-mediated lamellipodial tension instead depends predominantly on the coupling of this C-terminal TLN ABS to the actomyosin-driven retrograde actin flow force. This lamellipodial tension is crucial for full integrin activation eventually determining integrin-dependent cell migration. In the integrin/ligand-independent condition the frictional membrane resistance participates to these processes. Integrin/ligand binding can also contribute but is not necessarily required.
journal_name
Eur J Cell Bioljournal_title
European journal of cell biologyauthors
Schulte C,Ferraris GM,Oldani A,Galluzzi M,Podestà A,Puricelli L,de Lorenzi V,Lenardi C,Milani P,Sidenius Ndoi
10.1016/j.ejcb.2015.10.002subject
Has Abstractpub_date
2016-01-01 00:00:00pages
1-14issue
1eissn
0171-9335issn
1618-1298pii
S0171-9335(15)30012-1journal_volume
95pub_type
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