Abstract:
:Adhesion to the extracellular matrix regulates numerous changes in the actin cytoskeleton by regulating the activity of the Rho family of small GTPases. Here, we report that adhesion and the associated changes in cell shape and cytoskeletal tension are all required for GTP-bound RhoA to activate its downstream effector, ROCK. Using an in vitro kinase assay for endogenous ROCK, we found that cells in suspension, attached on substrates coated with low density fibronectin, or on spreading-restrictive micropatterned islands all exhibited low ROCK activity and correspondingly low myosin light chain phosphorylation, in the face of high levels of GTP-bound RhoA. In contrast, allowing cells to spread against substrates rescued ROCK and myosin activity. Interestingly, inhibition of tension with cytochalasin D or blebbistatin also inhibited ROCK activity within 20 min. The abrogation of ROCK activity by cell detachment or inhibition of tension could not be rescued by constitutively active RhoA-V14. These results suggest the existence of a feedback loop between cytoskeletal tension, adhesion maturation, and ROCK signaling that likely contributes to numerous mechanochemical processes.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Bhadriraju K,Yang M,Alom Ruiz S,Pirone D,Tan J,Chen CSdoi
10.1016/j.yexcr.2007.07.002subject
Has Abstractpub_date
2007-10-01 00:00:00pages
3616-23issue
16eissn
0014-4827issn
1090-2422pii
S0014-4827(07)00322-9journal_volume
313pub_type
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