Kinetoplastid kinetochore proteins KKT2 and KKT3 have unique centromere localization domains

The kinetochore is the macromolecular protein complex that assembles onto centromeric DNA and binds spindle microtubules. Evolutionarily divergent kinetoplastids have an unconventional set of kinetochore proteins. It remains unknown how kinetochores assemble at centromeres in these organisms. Here, we characterize KKT2 and KKT3 in the kinetoplastid parasite Trypanosoma brucei. In addition to the N-terminal kinase domain and C-terminal divergent polo boxes, these proteins have a central domain of unknown function. We show that KKT2 and KKT3 are important for the localization of several kinetochore proteins and that their central domains are sufficient for centromere localization. Crystal structures of the KKT2 central domain from two divergent kinetoplastids reveal a unique zinc-binding domain (termed the CL domain for centromere localization), which promotes its kinetochore localization in T. brucei. Mutations in the equivalent domain in KKT3 abolish its kinetochore localization and function. Our work shows that the unique central domains play a critical role in mediating the centromere localization of KKT2 and KKT3.

Automated summary

The study characterizes the KKT2 and KKT3 proteins in the kinetoplastid parasite Trypanosoma brucei, showing that they are important for the localization of various kinetochore proteins, with their central domains playing a critical role. Crystal structures of the KKT2 central domain reveal a unique zinc-binding domain that promotes its kinetochore localization, while mutations in the equivalent domain in KKT3 abolish its function. This work demonstrates the importance of the unique central domains in mediating the centromere localization of KKT2 and KKT3.