PXD072662

PXD072662 is an original dataset announced via ProteomeXchange.

Title	Assembly and phospho-regulatory mechanisms of the budding yeast outer kinetochore KMN complex
Description	Crosslinking of KMN outer kinetochore complex. During mitosis and meiosis kinetochores mediate interactions between chromosomes and spindle microtubules. Kinetochores are multi-megadalton protein complexes essential for chromosome segregation in all eukaryotes, however recent structural, functional, and evolutionary studies have revealed divergent mechanisms of kinetochore assembly. In this study, we use cryo-EM to understand the structural mechanisms by which the budding yeast microtubule-binding outer kinetochore KMN complex assembles, and how its interactions with the centromere-binding inner kinetochore are regulated. The ten-subunit KMN complex comprises three subcomplexes: Knl1c, Mis12cMtw1c and Ndc80c. We show that α-helical motifs in the C-termini of the Mis12cMtw1c subunits Dsn1, Mis12Mtw1 and Pmf1Nnf1 bind Knl1c and Ndc80c. At the opposite end of the Mis12cMtw1c stalk, an N-terminal auto-inhibitory segment of Dsn1 (Dsn1AI) folds into two α-helices that engage its head 1 domain, thereby occluding binding sites for the inner kinetochore subunits CENP-CMif2 and CENP-UAme1, reducing their affinity for Mis12cMtw1. Our structure reveals how Aurora BIpl1 phosphorylation of Dsn1AI would release this auto-inhibition to substantially strengthen pre-existing connections between the inner and outer kinetochore.
HostingRepository	PRIDE
AnnounceDate	2026-03-16
AnnouncementXML	Submission_2026-03-16_08:38:35.802.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Tomos Morgan
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: NEWT:4932;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2026-01-05 11:17:36	ID requested
⏵ 1	2026-03-16 08:38:36	announced

Dataset with its publication pending

submitter keyword: Kinetochore, CL-MS, Crosslinking

David Barford
contact affiliation	MRC - Laboratory of Molecular Biology
contact email	dbarford@mrc-lmb.cam.ac.uk
lab head
Tomos Morgan
contact affiliation	MRC-LMB
contact email	tmorgan@mrc-lmb.cam.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD072662
     1. Label: PRIDE project
     2. Name: Assembly and phospho-regulatory mechanisms of the budding yeast outer kinetochore KMN complex