We also knocked out individual doublet microtubule subunits, revealing which structural elements are important for movement. Together, our work provides a new framework for understanding how diverse molecular “rowers” coordinate ciliary motility.

Next, we systematically deleted each dynein gene and analyzed how these knockouts altered flagellar movement. The results were surprising: each dynein distinctly impacted motility, but not necessarily in the ways predicted from earlier studies.

Using Leishmania as a model, we determined the cryo-EM structure of the doublet microtubule to pinpoint the position of each dynein. This gave us a detailed map of where every “rower” sits on the ciliary “boat.”

In an eight-person rowing boat, each rower contributes to movement but also has a unique role: balancing, powering, or setting the rhythm and pace. In our latest collaborative work we asked – do the eight dynein “rowers” in #cilia and #flagella operate in the same way?

We also observe trypanosomatid-specific axoneme specializations. One example is the B-tubule ponticulus structure, which was first observed nearly 60 years ago! We find that the lumen-spanning structure is made up of three components, whose periodicity is established by a filamentous MIP.

Using CRISPR, we knocked out each of our identified proteins and tested the mutant swimming speed. Our analysis found that the doublet is surprisingly resilient to individual MIP knockout. However, we show that the evolutionarily conserved inner junction is uniquely sensitive to knockout.

Our structure revealed a highly specialized doublet containing 51 microtubule inner proteins (MIPs). Once resolved, we used Leishmania as a model to test the contribution of each MIP to motility.