The COMA complex interacts with Cse4 and positions Sli15/Ipl1 at the budding yeast inner kinetochore

Elife. 2019 May 21;8:e42879. doi: 10.7554/eLife.42879.

Abstract

Kinetochores are macromolecular protein complexes at centromeres that ensure accurate chromosome segregation by attaching chromosomes to spindle microtubules and integrating safeguard mechanisms. The inner kinetochore is assembled on CENP-A nucleosomes and has been implicated in establishing a kinetochore-associated pool of Aurora B kinase, a chromosomal passenger complex (CPC) subunit, which is essential for chromosome biorientation. By performing crosslink-guided in vitro reconstitution of budding yeast kinetochore complexes we showed that the Ame1/Okp1CENP-U/Q heterodimer, which forms the COMA complex with Ctf19/Mcm21CENP-P/O, selectively bound Cse4CENP-A nucleosomes through the Cse4 N-terminus. The Sli15/Ipl1INCENP/Aurora-B core-CPC interacted with COMA in vitro through the Ctf19 C-terminus whose deletion affected chromosome segregation fidelity in Sli15 wild-type cells. Tethering Sli15 to Ame1/Okp1 rescued synthetic lethality upon Ctf19 depletion in a Sli15 centromere-targeting deficient mutant. This study shows molecular characteristics of the point-centromere kinetochore architecture and suggests a role for the Ctf19 C-terminus in mediating CPC-binding and accurate chromosome segregation.

Keywords: S. cerevisiae; aurora B; biochemistry; cell biology; chemical biology; chemical crosslinking; chromosomal passenger complex; chromosome segregation; error correction; feedback control; kinetochore; mass spectrometry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Kinetochores / chemistry*
  • Protein Binding
  • Protein Interaction Maps*
  • Saccharomyces cerevisiae Proteins / analysis*
  • Saccharomycetales / chemistry*

Substances

  • Saccharomyces cerevisiae Proteins