A new function for adducin. Calcium/calmodulin-regulated capping of the barbed ends of actin filaments.
Adducin is a membrane skeleton protein originally described in human erythrocytes that promotes the binding of spectrin to actin and also binds directly to actin and bundles actin filaments. Adducin is associated with regions of cell-cell contact in nonerythroid cells, where it is believed to play a role in regulating the assembly of the spectrin-actin membrane skeleton. In this study we demonstrate a novel function for adducin; it completely blocks elongation and depolymerization at the barbed (fast growing) ends of actin filaments, thus functioning as a barbed end capping protein (Kcap approximately 100 nM). This barbed end capping activity requires the intact adducin molecule and is not provided by the NH2-terminal globular head domains alone nor by the COOH-terminal extended tail domains, which were previously shown to contain the spectrin-actin binding, calmodulin binding, and phosphorylation sites. A novel difference between adducin and other previously described capping proteins is that it is down-regulated by calmodulin in the presence of calcium. The association of stoichiometric amounts of adducin with the short erythrocyte actin filaments in the membrane skeleton indicates that adducin could be the functional barbed end capper in erythrocytes and play a role in restricting actin filament length. Our experiments also suggest novel possibilities for calcium regulation of actin filament assembly by adducin in erythrocytes and at cell-cell contact sites in nonerythroid cells.
Kuhlman
P A
PA
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Hughes
C A
CA
Bennett
V
V
Fowler
V M
VM
D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000502:physiology; D002148:Calmodulin-Binding Proteins / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D013049:Spectrin / Q000378:metabolism; D000099291:Adducins
D000199:Actins; D002118:Calcium; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D003598:Cytoskeletal Proteins; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D013049:Spectrin; D000099291:Adducins