Caldesmon inhibits nonmuscle cell contractility and interferes with the formation of focal adhesions.
Caldesmon is known to inhibit the ATPase activity of actomyosin in a Ca(2+)-calmodulin-regulated manner. Although a nonmuscle isoform of caldesmon is widely expressed, its functional role has not yet been elucidated. We studied the effects of nonmuscle caldesmon on cellular contractility, actin cytoskeletal organization, and the formation of focal adhesions in fibroblasts. Transient transfection of nonmuscle caldesmon prevents myosin II-dependent cell contractility and induces a decrease in the number and size of tyrosine-phosphorylated focal adhesions. Expression of caldesmon interferes with Rho A-V14-mediated formation of focal adhesions and stress fibers as well as with formation of focal adhesions induced by microtubule disruption. This inhibitory effect depends on the actin- and myosin-binding regions of caldesmon, because a truncated variant lacking both of these regions is inactive. The effects of caldesmon are blocked by the ionophore A23187, thapsigargin, and membrane depolarization, presumably because of the ability of Ca(2+)-calmodulin or Ca(2+)-S100 proteins to antagonize the inhibitory function of caldesmon on actomyosin contraction. These results indicate a role for nonmuscle caldesmon in the physiological regulation of actomyosin contractility and adhesion-dependent signaling and further demonstrate the involvement of contractility in focal adhesion formation.
Helfman
D M
DM
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
Levy
E T
ET
Berthier
C
C
Shtutman
M
M
Riveline
D
D
Grosheva
I
I
Lachish-Zalait
A
A
Elbaum
M
M
Bershadsky
A D
AD
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000205:Actomyosin / Q000378:metabolism; D000818:Animals; D001905:Botulinum Toxins / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002461:Cell Line, Transformed; D003599:Cytoskeleton / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D009154:Mutation; D015739:Nocodazole / Q000494:pharmacology; D051381:Rats; D019284:Thapsigargin / Q000494:pharmacology; D014162:Transfection
D002135:Calcium-Binding Proteins; D002148:Calmodulin-Binding Proteins; D004791:Enzyme Inhibitors; D007476:Ionophores; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D000001:Calcimycin; D019284:Thapsigargin; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000205:Actomyosin; D001905:Botulinum Toxins; D015739:Nocodazole