Structure of the ciliary tip central pair reveals the unique role of the microtubule-seam binding protein SPEF1

Legal, T.; Joachimiak, E.; Parra, M.; Peng, W.; Tam, A.; Black, C. S.; Valente-Paterno, M.; Brouhard, G.; Gaertig, J.; Wloga, D.; Bui, K. H.

doi:10.1101/2024.12.02.626492

bioRxiv · 10.1101/2024.12.02.626492

Structure of the ciliary tip central pair reveals the unique role of the microtubule-seam binding protein SPEF1

Legal, T.·Joachimiak, E.·Parra, M.·Peng, W.·Tam, A.·Black, C. S.·Valente-Paterno, M.·Brouhard, G.·Gaertig, J.·Wloga, D.·Bui, K. H.

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Abstract

Motile cilia are unique organelles with the ability to autonomously move. Force generated by beating cilia propels cells and moves fluids. The ciliary skeleton is made of peripheral doublet microtubules and a central pair (CP) with a distinct structure at the tip. In this study, we present a high-resolution structure of the CP in the ciliary tip of the ciliate Tetrahymena thermophila and identify several tip proteins that bind and form unique patterns on both microtubules of the tip CP. Two of those proteins that contain tubulin polymerization-promoting protein (TPPP)-like domains, TLP1 and TLP2, bind to high curvature regions of the microtubule. TLP2, which contains two TPPP-like domains, is an unusually long protein that wraps laterally around half a microtubule and forms the bridge between the two microtubules. Moreover, we found that the conserved protein SPEF1 binds to both microtubule seams. In vitro, human SPEF1 not only binds to the microtubule seam but also crosslinks two parallel microtubules. Single-molecule microtubule dynamics assays indicate that SPEF1 stabilizes microtubules in vitro. Together, these data show that the proteins in the tip CP maintain stable microtubule structure and probably play important roles in maintaining the integrity of the axoneme.

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Legal, T., Joachimiak, E., Parra, M., Peng, W., Tam, A., Black, C. S., Valente-Paterno, M., Brouhard, G., Gaertig, J., Wloga, D., Bui, K. H.. 2024-12-04. Structure of the ciliary tip central pair reveals the unique role of the microtubule-seam binding protein SPEF1. https://doi.org/10.1101/2024.12.02.626492

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