Hsp90–Sgt1 and Skp1 target human Mis12 complexes to ensure efficient formation of kinetochore–microtubule binding sites

Abstract

The Hsp90–Sgt1 chaperone and the ubiquitin ligase subunit Skp1 regulate the assembly and turnover of the kinetochore complex Mis12.

Abstract

The formation of functional kinetochores requires the accurate assembly of a large number of protein complexes. The Hsp90–Sgt1 chaperone complex is important for this process; however, its targets are not conserved and its exact contribution to kinetochore assembly is unclear. Here, we show that human Hsp90–Sgt1 interacts with the Mis12 complex, a so-called keystone complex required to assemble a large fraction of the kinetochore. Inhibition of Hsp90 or Sgt1 destabilizes the Mis12 complex and delays proper chromosome alignment due to inefficient formation of microtubule-binding sites. Interestingly, coinhibition of Sgt1 and the SCF subunit, Skp1, increases Mis12 complexes at kinetochores and restores timely chromosome alignment but forms less-robust microtubule-binding sites. We propose that a balance of Mis12 complex assembly and turnover is required for the efficient and accurate assembly of kinetochore–microtubule binding sites. These findings support a novel role for Hsp90–Sgt1 chaperones in ensuring the fidelity of multiprotein complex assembly.