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Science 3 November 1989:
Vol. 246. no. 4930, pp. 656 - 658
DOI: 10.1126/science.2530629
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Articles

Science, Vol 246, Issue 4930, 656-658
Copyright © 1989 by American Association for the Advancement of Science

articles

Expression and characterization of a functional myosin head fragment in Dictyostelium discoideum

DJ Manstein, KM Ruppel, and JA Spudich

Department of Cell Biology, Stanford University School of Medicine, CA 94305.

The isolated head fragment of myosin is a motor protein that is able to use energy liberated from the hydrolysis of adenosine triphosphate to cause sliding movement of actin filaments. Expression of a myosin fragment nearly equivalent to the amino-terminal globular head domain, generally referred to as subfragment 1, has been achieved by transforming the eukaryotic organism Dictyostelium discoideum with a plasmid that carries a 2.6-kilobase fragment of the cloned Dictyostelium myosin heavy chain gene under the control of the Dictyostelium actin-15 promoter. The recombinant fragment of the myosin heavy chain was purified 2400-fold from one of the resulting cell lines and was found to be functional by the following criteria: the myosin head fragment copurified with the essential and regulatory myosin light chains, decorated actin filaments, and displayed actin-activated adenosine triphosphatase activity. In addition, motility assays in vitro showed that the recombinant myosin fragment is capable of supporting sliding movement of actin filaments.


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A cell number-counting factor regulates the cytoskeleton and cell motility in Dictyostelium.
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