A dynein arm attachment cycle produces sliding between adjacent doublet microtubules (N and N + 1) of cilia. In intact axonemes, in the absence of ATP, almost all arms appear attached at both ends (rigor). When ATP is added, most arms detach from doublet N + 1. In ATP and vanadate, the arms do not return to rigor, suggesting that ATP hydrolysis is required for re-extension and reattachment of the dynein arm, but not for detachment. Using solutions containing dynein to decorate dynein-less axonemal doublets, we confirm this interpretation. In the absence of ATP, both sides of each doublet decorate with arms. Addition of ATP, ATP and vanadate or AMP-PNP causes immediate arm detachment, but only in the first instance, where extensive ATP hydrolysis can occur, does decoration eventually reappear. Dynein decorates heterologous axonemal doublets and brain microtubules, as well as homologous doublets, suggesting that this mechanochemical cycle may have general applicability in microtubule-based cell motility.