title: The molecular architecture of the Dam1 kinetochore complex is defined by cross-linking based structural modelling.
abstract: Accurate segregation of chromosomes during cell division is essential. The Dam1 complex binds kinetochores to microtubules and its oligomerization is required to form strong attachments. It is a key target of Aurora B kinase, which destabilizes erroneous attachments allowing subsequent correction. Understanding the roles and regulation of the Dam1 complex requires structural information. Here we apply cross-linking/mass spectrometry and structural modelling to determine the molecular architecture of the Dam1 complex. We find microtubule attachment is accompanied by substantial conformational changes, with direct binding mediated by the carboxy termini of Dam1p and Duo1p. Aurora B phosphorylation of Dam1p C terminus weakens direct interaction with the microtubule. Furthermore, the Dam1p amino terminus forms an interaction interface between Dam1 complexes, which is also disrupted by phosphorylation. Our results demonstrate that Aurora B inhibits both direct interaction with the microtubule and oligomerization of the Dam1 complex to drive error correction during mitosis.
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Alex
authors.initials: A
authors.lastname: Zelter
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bonomi
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Jae Ook
authors.initials: JO
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Neil T
authors.initials: NT
authors.lastname: Umbreit
authors.affiliation: Institute for Systems Biology, Seattle, Washington 98109, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Hoopmann
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Johnson
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Riffle
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Jaschob
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: MacCoss
authors.affiliation: Institute for Systems Biology, Seattle, Washington 98109, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Moritz
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Trisha N
authors.identifier: http://orcid.org/0000-0003-4797-3152
authors.initials: TN
authors.lastname: Davis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D064107:Aurora Kinase B / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D006801:Humans; D018386:Kinetochores / Q000737:chemistry / Q000502:physiology*; D013058:Mass Spectrometry; D008870:Microtubules; D008958:Models, Molecular; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: C120624:BCAS2 protein, human; D009363:Neoplasm Proteins; C576921:AURKB protein, human; D064107:Aurora Kinase B