Molecular organization of the distal tip of vertebrate motile cilia

The beating of cilia on multi-ciliated cells (MCCs) is essential for normal development and homeostasis in animals. Unlike basal bodies or axonemes, the distal tips of MCC cilia remain poorly defined. Here, we characterize the molecular organization of the distal tip of vertebrate MCC cilia, revealing two distinct domains occupied by distinct protein constituents. Using frog, mouse, and human MCCs, we find that two largely uncharacterized proteins, Ccdc78 and Ccdc33 occupy a specialized region at the extreme distal tip, and these are required for the normal organization of other tip proteins, including Spef1, Cep104, and Eb3. Ccdc78 and Cccdc33 are also independently required for normal length regulation of MCC cilia. Mechanistically, Ccdc78 and Ccdc33 display robust microtubule-bundling activity both in vivo and in vitro . Thus, we reveal that two previously undefined proteins form a key module for organizing and stabilizing the distal tip of motile cilia in vertebrate MCC. We propose that these proteins represent potential disease loci for motile ciliopathies.