DirectDemonstrationofActinFilamentAnnealingInVitro.PDFVIP

Published June 1, 1988 Direct Demonstration of Actin Filament Annealing In Vitro Douglas B. Murphy, Rafael O. Gray, William A. Grasser, and T h o m a s D. Pollard Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 Abstract. Direct electron microscopic examination collision of two filament ends with an apparent anneal- confirms that short actin filaments rapidly anneal end- ing rate constant of 107 M-Is -I. This rapid phase lasts to-end in vitro, leading over time to an increase in 10 s and is followed by a slow phase lasting minutes filament length at steady state. During annealing of to hours. Annealing is calculated to contribute mini- mixtures of native unlabeled filaments and glutaralde- mally to filament elongation during the initial stages of hyde-fixed filaments labeled with myosin subfragment-1, self-assembly. However, the rapid rate of annealing of the structural polarity within heteropolymers is con- sonicated fixed filaments observed in vitro suggests served absolutely. Annealing does not appear to re- that it may be an efficient mechanism for repairing D quire either ATP hydrolysis or the presence of exoge- breaks in filaments and that annealing together with o w nous actin monomers, sugg