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Published 9 July 2001. doi:10.1083/jcb.200011013
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© The Rockefeller University Press, 0021-9525/2001/7/137 $5.00
The Journal of Cell Biology, Volume 154, Number 1, July 9, 2001 137-146

Article

Myosin II dynamics and cortical flow during contractile ring formation in Dictyostelium cells



Shigehiko Yumura

Department of Biology, Faculty of Science, Yamaguchi University, Yamaguchi 753-8512, Japan

Address correspondence to Shigehiko Yumura, Department of Biology, Faculty of Science, Yamaguchi University, Yoshida 1677-1, Yamaguchi 753-8512, Japan. Tel.: 81-83-933-5717. Fax: 81-83-933-5768. E-mail: yumura{at}po.cc.yamaguchi-u.ac.jp

Myosin II is a major component of a contractile ring. To examine if myosin II turns over in contractile rings, fluorescence of GFP–myosin II expressed in Dictyostelium cells was bleached locally by laser illumination, and the recovery was monitored. The fluorescence recovered with a half time of 7.01 ± 2.62 s. This recovery was not caused by lateral movement of myosin II from the nonbleached area, but by an exchange with endoplasmic myosin II. Similar experiments were performed in cells expressing GFP–3ALA myosin II, of which three phosphorylatable threonine residues were replaced with alanine residues. In this case, recovery was not detected within a comparable time range. These results indicate that myosin II in the contractile ring performs dynamic turnover via its heavy chain phosphorylation. Because GFP–3ALA myosin II did not show the recovery, it served as a useful marker of myosin II movement, which enabled us to demonstrate cortical flow of myosin II toward the equator for the first time. Thus, cortical flow accompanies the dynamic exchange of myosin II during the formation of contractile rings.

Key Words: FRAP; cleavage furrow; myosin II; cortical flow; GFP


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