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J Cell Sci. 2016 Apr 15;129(8):1547-51. doi: 10.1242/jcs.184598. Epub 2016 Mar 1.

Docking-complex-independent alignment of Chlamydomonas outer dynein arms with 24-nm periodicity in vitro.

Author information

  • 1Department of Cell Biology and Anatomy, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan Department of Anatomy and Structural Biology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokatoh, Chuo, Yamanashi 409-3898, Japan toda@yamanashi.ac.jp.
  • 2Department of Cell Biology and Anatomy, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Abstract

The docking complex is a molecular complex necessary for assembly of outer dynein arms (ODAs) on the axonemal doublet microtubules (DMTs) in cilia and flagella. The docking complex is hypothesized to be a 24-nm molecular ruler because ODAs align along the DMTs with 24-nm periodicity. In this study, we rigorously tested this hypothesis using structural and genetic methods. We found that the ODAs can bind to DMTs and porcine microtubules with 24-nm periodicities even in the absence of the docking complexin vitro Using cryo-electron tomography and structural labeling, we observed that the docking complex took an unexpectedly flexible conformation and did not lie along the length of DMTs. In the absence of docking complex, ODAs were released from the DMT at relatively low ionic strength conditions, suggesting that the docking complex strengthens the electrostatic interactions between the ODA and DMT. Based on these results, we conclude that the docking complex serves as a flexible stabilizer of the ODA rather than as a molecular ruler.

KEYWORDS:

Axoneme; Cilia; Docking complex; Flagella; Microtubule; Outer dynein arm

PMID:
26933181
DOI:
10.1242/jcs.184598
[PubMed - indexed for MEDLINE]
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