Regulation of actin binding and actin bundling activities of fascin by caldesmon coupled with tropomyosin.
Human fascin is an actin-bundling protein and is thought to play a role in the formation of microfilament bundles of microspikes and stress fibers in cultured cells. To explore the regulation of fascin-actin interaction, we have examined the effects of culture cell caldesmon and tropomyosin (TM) on actin binding activity of human fascin. Caldesmon alone or TM alone has little or no effect on the actin binding of fascin. However, caldesmon together with TM completely inhibits actin binding of human fascin. When calmodulin is added, the inhibition of fascin-actin interaction by caldesmon and TM becomes Ca2+ dependent because Ca2+/calmodulin blocks actin binding of caldesmon. Furthermore, as phosphorylation of caldesmon by cdc2 kinase inhibits actin binding of caldesmon, phosphorylation can also control actin binding of fascin in the presence of TM. As expected by the inhibition of fascin-actin binding, caldesmon coupled with TM also inhibits actin bundling activity of fascin. Whereas smooth muscle caldesmon alone or TM alone shows no effect, caldesmon together with TM completely inhibits actin bundling activity of fascin. This inhibition is again Ca2+ dependent when calmodulin is added to the system. These results suggest important roles for caldesmon and TM in the regulation of the function of human fascin.
Department of Molecular Biology and Biochemistry, Nelson Laboratory, Busch Campus, Rutgers University, Piscataway, New Jersey 08854, USA.
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R
Ishikawa
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S
Yamashiro
K
K
Kohama
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F
Matsumura
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002461:Cell Line, Transformed; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010766:Phosphorylation; D011485:Protein Binding; D051381:Rats; D014335:Tropomyosin / Q000378:metabolism*
D000199:Actins; D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins; D008840:Microfilament Proteins; D014335:Tropomyosin; C020324:fascin