Structure of the human inner kinetochore bound to a centromeric CENP-A nucleosome

Kinetochores assemble onto specialized centromeric CENP-A (centromere protein A) nucleosomes (CENP-A(Nuc)) to mediate attachments between chromosomes and the mitotic spindle. We describe cryo-electron microscopy structures of the human inner kinetochore constitutive centromere associated network (CCAN) complex bound to CENP-A(Nuc) reconstituted onto alpha-satellite DNA. CCAN forms edge-on contacts with CENP-A(Nuc), and a linker DNA segment of the alpha-satellite repeat emerges from the fully wrapped end of the nucleosome to thread through the central CENP-LN channel that tightly grips the DNA. The CENP-TWSX histone-fold module further augments DNA binding and partially wraps the linker DNA in a manner reminiscent of canonical nucleosomes. Our study suggests that the topological entrapment of the linker DNA by CCAN provides a robust mechanism by which kinetochores withstand both pushing and pulling forces exerted by the mitotic spindle.