Cyclase-associated protein (CAP) acts directly on F-actin to accelerate cofilin-mediated actin severing across the range of physiological pH.
Fast actin depolymerization is necessary for cells to rapidly reorganize actin filament networks. Utilizing a Listeria fluorescent actin comet tail assay to monitor actin disassembly rates, we observed that although a mixture of actin disassembly factors (cofilin, coronin, and actin-interacting protein 1 is sufficient to disassemble actin comet tails in the presence of physiological G-actin concentrations this mixture was insufficient to disassemble actin comet tails in the presence of physiological F-actin concentrations. Using biochemical complementation, we purified cyclase-associated protein (CAP) from thymus extracts as a factor that protects against the inhibition of excess F-actin. CAP has been shown to participate in actin dynamics but has been thought to act by liberating cofilin from ADP·G-actin monomers to restore cofilin activity. However, we found that CAP augments cofilin-mediated disassembly by accelerating the rate of cofilin-mediated severing. We also demonstrated that CAP acts directly on F-actin and severs actin filaments at acidic, but not neutral, pH. At the neutral pH characteristic of cytosol in most mammalian cells, we demonstrated that neither CAP nor cofilin are capable of severing actin filaments. However, the combination of CAP and cofilin rapidly severed actin at all pH values across the physiological range. Therefore, our results reveal a new function for CAP in accelerating cofilin-mediated actin filament severing and provide a mechanism through which cells can maintain high actin turnover rates without having to alkalinize cytosol, which would affect many biochemical reactions beyond actin depolymerization.
Department of Molecular and Integrative Physiology, University of Illinois, Urbana, Illinois 61801.
Kieran P M
KPM
Normoyle
Department of Cell and Developmental Biology, University of Illinois, Urbana, Illinois 61801. Electronic address: wbrieher@illinois.edu.
William M
WM
Brieher
D008841:Actin Cytoskeleton / Q000737:chemistry*; D051339:Actin Depolymerizing Factors / Q000737:chemistry*; D000199:Actins / Q000737:chemistry*; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry*; D002417:Cattle; D006801:Humans; D006863:Hydrogen-Ion Concentration
D051339:Actin Depolymerizing Factors; D000199:Actins; D002352:Carrier Proteins