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

Abstract

The docking complex (DC) is a molecular complex necessary for assembly of outer dynein arms (ODAs) on the axonemal doublet microtubules (DMTs) in cilia and flagella. The DC 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 could bind to DMTs and porcine microtubules with 24-nm periodicities even in the absence of DC in vitro. Using cryo-electron tomography and structural labeling, we observed that the DC took an unexpectedly flexible conformation and did not lie along the length of DMTs. In the absence of DC, ODAs were released from the DMT at relatively low ionic strength, suggesting that DC strengthens the electrostatic interactions between the ODA and DMT. Based on these results, we concluded that the DC serves as a flexible stabilizer of the ODA rather than a molecular ruler.