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Published online October 13, 2008
doi:10.1083/jcb.200802025
The Journal of Cell Biology
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Ahmed et al.
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ARTICLE

ODA16 aids axonemal outer row dynein assembly through an interaction with the intraflagellar transport machinery

Noveera T. Ahmed1, Chunlei Gao1, Ben F. Lucker2, Douglas G. Cole2, and David R. Mitchell1

1 Department of Cell and Developmental Biology, State University of New York Upstate Medical University, Syracuse, NY 13210
2 Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83844

Correspondence to David R. Mitchell: mitcheld{at}upstate.edu

Formation of flagellar outer dynein arms in Chlamydomonas reinhardtii requires the ODA16 protein at a previously uncharacterized assembly step. Here, we show that dynein extracted from wild-type axonemes can rebind to oda16 axonemes in vitro, and dynein in oda16 cytoplasmic extracts can bind to docking sites on pf28 (oda) axonemes, which is consistent with a role for ODA16 in dynein transport, rather than subunit preassembly or binding site formation. ODA16 localization resembles that seen for intraflagellar transport (IFT) proteins, and flagellar abundance of ODA16 depends on IFT. Yeast two-hybrid analysis with mammalian homologues identified an IFT complex B subunit, IFT46, as a directly interacting partner of ODA16. Interaction between Chlamydomonas ODA16 and IFT46 was confirmed through in vitro pull-down assays and coimmunoprecipitation from flagellar extracts. ODA16 appears to function as a cargo-specific adaptor between IFT particles and outer row dynein needed for efficient dynein transport into the flagellar compartment.

Abbreviations used in this paper: IFT, intraflagellar transport; ODA-HC and ODA-IC, outer arm dynein heavy chain and intermediate chain, respectively.

© 2008 Ahmed et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).


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