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Published online April 16, 2007
doi:10.1083/jcb.200609038
The Journal of Cell Biology, Vol. 177, No. 2, 243-252
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Oda et al.
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Article

Three-dimensional structures of the flagellar dynein–microtubule complex by cryoelectron microscopy



Toshiyuki Oda1,2, Nobutaka Hirokawa2, and Masahide Kikkawa1

1 Department of Cell Biology, University of Texas, Southwestern Medical Center, Dallas, TX 75390
2 Department of Cell Biology and Anatomy, Graduate School of Medicine, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan

Correspondence to Masahide Kikkawa: mkikkawa{at}gmail.com

The outer dynein arms (ODAs) of the flagellar axoneme generate forces needed for flagellar beating. Elucidation of the mechanisms underlying the chemomechanical energy conversion by the dynein arms and their orchestrated movement in cilia/flagella is of great importance, but the nucleotide-dependent three-dimensional (3D) movement of dynein has not yet been observed. In this study, we establish a new method for reconstructing the 3D structure of the in vitro reconstituted ODA–microtubule complex and visualize nucleotide-dependent conformational changes using cryoelectron microscopy and image analysis. As the complex went from the rigor state to the relaxed state, the head domain of the ß heavy chain shifted by 3.7 nm toward the B tubule and inclined 44° inwards. These observations suggest that there is a mechanism that converts head movement into the axonemal sliding motion.

Abbreviations used in this paper: MT, microtubule; ODA, outer dynein arm; ODA–CB-MT, ODA–cross-bridged MT; QFDE, quick freeze deep etch.


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