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

Abstract

Formation of flagellar outer dynein arms in Chlam-ydomonas reinhardtii requires the ODA 16 protein at a previously uncharacterized assembly step. Here, we show that dynein extracted from wild-type ax-onemes 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 homo-logues identified an IFT complex B subunit, IFT46, as a directly interacting partner of ODA 16. 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. © 2008 Ahmed et al.