Architecture and flexibility of native kinetochores revealed by structural studies utilizing a thermophilic yeast

janelia7_blocks-janelia7_biblio_header | block

bioRxiv. 2024 Mar 1;. doi: 10.1101/2024.02.28.582571

Architecture and flexibility of native kinetochores revealed by structural studies utilizing a thermophilic yeast Cryo-Electron Microscopy

Barrero Daniel, Wijeratne Sithara, Zhao Xiaowei, Cunningham Grace, Yan Rui, Nelson Christian, Arimura Yasuhiro, Funabiki Hironori, Asbury Charles, Yu Zhiheng, Subramanian Radhika, Biggins Sue

janelia7_blocks-janelia7_biblio_abstract | block

Abstract

Eukaryotic chromosome segregation requires kinetochores, multi-megadalton protein machines that assemble on the centromeres of chromosomes and mediate attachments to dynamic spindle microtubules. Kinetochores are built from numerous complexes, and understanding how they are arranged is key to understanding how kinetochores perform their multiple functions. However, an integrated understanding of kinetochore architecture has not yet been established. To address this, we purified functional, native kinetochores from Kluyveromyces marxianus and examined them by electron microscopy, cryo-electron tomography and atomic force microscopy. The kinetochores are extremely large, flexible assemblies that exhibit features consistent with prior models. We assigned kinetochore polarity by visualizing their interactions with microtubules and locating the microtubule binder Ndc80c. This work shows that isolated kinetochores are more dynamic and complex than what might be anticipated based on the known structures of recombinant subassemblies, and provides the foundation to study the global architecture and functions of kinetochores at a structural level.

node:body | entity_field

bioRxiv preprint https://doi.org/10.1101/2024.02.28.582571

Main Menu (Mobile)- Block

Main Menu - Block

Architecture and flexibility of native kinetochores revealed by structural studies utilizing a thermophilic yeast

Abstract

Janelia Authors