The Role of Tubulin Polyglycylation and Polyglutamylation in Ciliary Mechanics

Tubulin post-translational modifications (tPTMs) are currently studied as vital, yet obscure, cytoskeletal regulators. Their regulatory function relies on the spatiotemporal control over the activity of multiple tubulin modifying enzymes that functionalize microtubules, enabling their differentiation. The cilium, one of the organelles with the richest tPTMs diversity, has been studied with determination for the last decades, allowing the interrogation of the molecular processes that give rise to its function. The inner structure of this thin organelle, the axoneme, comprises a microtubule scaffold periodically decorated with macromolecular complexes whose characterization has been achieved with pseudoatomic detail. The molecular distribution and mechanism of action of tPTMs in cilia remain elusive. Using a combination of immunolabelling and cryoelectron tomography we interrogated the molecular function two tPTMs in the axonemal context. We showed that tubulin polyglycylation spanned most of the microtubular surface and was required for axonemal dynein activity regulation and male fertility. Additionally, there was an enrichment of polyglutamylation on a single microtubule protofilament, forming a pattern complementary to that of polyglycylation, that was required for proper coupling of microtubule sliding and bending forces during ciliary beating