TY - JOUR
T1 - Spatial distribution and functional significance of activated vinculin in living cells
AU - Chen, Hui
AU - Cohen, Daniel M.
AU - Choudhury, Dilshad M.
AU - Kioka, Noriyuki
AU - Craig, Susan W.
PY - 2005/5/9
Y1 - 2005/5/9
N2 - Conformational change is believed to be important to vinculin's function at sites of cell adhesion. However, nothing is known about vinculin's conformation in living cells. Using a Forster resonance energy transfer probe that reports on changes in vinculin's conformation, we find that vinculin is in the actin-binding conformation in a peripheral band of adhesive puncta in spreading cells. However, in fully spread cells with established polarity, vinculin's conformation is variable at focal adhesions. Time-lapse imaging reveals a gradient of conformational change that precedes loss of vinculin from focal adhesions in retracting regions. At stable or protruding regions, recruitment of vinculin is not necessarily coupled to the actin-binding conformation. However, a different measure of vinculin conformation, the recruitment of vinexin β by activated vinculin, shows that autoinhibition of endogenous vinculin is relaxed at focal adhesions. Beyond providing direct evidence that vinculin is activated at focal adhesions, this study shows that the specific functional conformation correlates with regional cellular dynamics.
AB - Conformational change is believed to be important to vinculin's function at sites of cell adhesion. However, nothing is known about vinculin's conformation in living cells. Using a Forster resonance energy transfer probe that reports on changes in vinculin's conformation, we find that vinculin is in the actin-binding conformation in a peripheral band of adhesive puncta in spreading cells. However, in fully spread cells with established polarity, vinculin's conformation is variable at focal adhesions. Time-lapse imaging reveals a gradient of conformational change that precedes loss of vinculin from focal adhesions in retracting regions. At stable or protruding regions, recruitment of vinculin is not necessarily coupled to the actin-binding conformation. However, a different measure of vinculin conformation, the recruitment of vinexin β by activated vinculin, shows that autoinhibition of endogenous vinculin is relaxed at focal adhesions. Beyond providing direct evidence that vinculin is activated at focal adhesions, this study shows that the specific functional conformation correlates with regional cellular dynamics.
UR - http://www.scopus.com/inward/record.url?scp=18844369343&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=18844369343&partnerID=8YFLogxK
U2 - 10.1083/jcb.200410100
DO - 10.1083/jcb.200410100
M3 - Article
C2 - 15883197
AN - SCOPUS:18844369343
SN - 0021-9525
VL - 169
SP - 459
EP - 470
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 3
ER -