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

ABSTRACT: Kinetochores form the link between chromosomes and microtubules of the mitotic spindle. The heterodecameric Dam1 complex (Dam1c) is a major component of the S. cerevisiae outer kinetochore, assembling into 3 MDa-sized microtubule-embracing rings, but how ring assembly is specifically initiated at kinetochores 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.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Franz Herzog

LAB HEAD: Franz Herzog

PROVIDER: PXD026935 | Pride | 2021-06-28

REPOSITORIES: Pride

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			Action	DRS
	AG_Herzog_rombaut_E1607_223.raw	Raw
	AG_Herzog_rombaut_E1607_223_matched.spec.xml	Xml
	AG_Herzog_rombaut_E1607_224.raw	Raw
	AG_Herzog_rombaut_E1607_224_matched.spec.xml	Xml
	AG_Herzog_rombaut_E1607_226.raw	Raw

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Mapping of cohesion factor association sites in S. cerevisiae - Meiosis I chromosome segregation is established through regulation of microtubule-kinetochore interactions.

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Shugoshin biases chromosomes for biorientation through condensin recruitment to the pericentromere

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