Phospho-regulated Bim1/EB1 interactions trigger Dam1c ring assembly at the budding yeast outer kinetochore

Alexander Dudziak, Lena Engelhard, Cole Bourque, Björn Udo Klink, Pascaline Rombaut, Nikolay Kornakov, Karolin Jänen, Franz Herzog, Christos Gatsogiannis, Stefan Westermann

Research output: Contribution to journalArticlepeer-review

Abstract

Kinetochores form the link between chromosomes and microtubules of the mitotic spindle. The heterodecameric Dam1 complex (Dam1c) is a major component of the Saccharomyces cerevisiae outer kinetochore, assembling into 3 MDa-sized microtubule-embracing rings, but how ring assembly is specifically initiated in vivo remains to be understood. Here, we describe a molecular pathway that provides local control of ring assembly during the establishment of sister kinetochore bi-orientation. We show that Dam1c and the general microtubule plus end-associated protein (+TIP) Bim1/EB1 form a stable complex depending on a conserved motif in the Duo1 subunit of Dam1c. EM analyses reveal that Bim1 crosslinks protrusion domains of adjacent Dam1c heterodecamers and promotes the formation of oligomers with defined curvature. Disruption of the Dam1c-Bim1 interaction impairs kinetochore localization of Dam1c in metaphase and delays mitosis. Phosphorylation promotes Dam1c-Bim1 binding by relieving an intramolecular inhibition of the Dam1 C-terminus. In addition, Bim1 recruits Bik1/CLIP-170 to Dam1c and induces formation of full rings even in the absence of microtubules. Our data help to explain how new kinetochore end-on attachments are formed during the process of attachment error correction.

Original languageEnglish
Article numbere108004
Pages (from-to)e108004
JournalEMBO Journal
Volume40
Issue number18
DOIs
Publication statusPublished - 15 Sept 2021
Externally publishedYes

Keywords

  • bi-orientation
  • Bik1
  • Bim1
  • chromosome segregation
  • Dam1/DASH complex
  • Kinetochores/metabolism
  • Mitosis/physiology
  • Chromosome Segregation
  • Phosphorylation
  • Spindle Apparatus/metabolism
  • Saccharomycetales/physiology
  • Multiprotein Complexes/metabolism
  • Microtubule Proteins/metabolism
  • Protein Binding
  • Microtubule-Associated Proteins/metabolism

Research fields

  • Cell Division
  • Chemical Crosslinking
  • Mass spectrometry

IMC Research Focuses

  • Medical biotechnology

ÖFOS 2012 - Austrian Fields of Study

  • 106037 Proteomics
  • 106041 Structural biology
  • 106044 Systems biology

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