PXD047133-2

PXD047133 is an original dataset announced via ProteomeXchange.

Title	Kinetochores read detyrosinated tubulin to sustain microtubule attachments
Description	Post-translational cycles of α-tubulin detyrosination/tyrosination generate microtubule diversity, with broad physiological and clinical implications. However, the mechanistic understanding of how this microtubule diversity is decoded into specific cellular functions remains largely unknown. Here we show that the core microtubule-binding site at kinetochores ‘reads’ and responds to the α-tubulin detyrosination/tyrosination code. By combining CRISPR-Cas9 gene editing of enzymatic ‘writers’ of α-tubulin detyrosination/tyrosination in human cells, with biochemical purifications, mass spectrometry and super-resolution microscopy, we identified over a hundred proteins that preferentially associate with either detyrosinated or tyrosinated microtubules during mitosis. Among these, the conserved KNL-1/MIS12 complex/NDC80 complex (KMN) network at kinetochores showed a preference for tyrosinated microtubules. In addition to established roles in error correction and polar chromosome alignment, live-cell imaging uncovered a previously overlooked link between the α-tubulin detyrosination/tyrosination code and the formation of functional kinetochore-microtubule attachments, necessary for normal metaphase chromosome oscillations and timely spindle assembly checkpoint satisfaction. In vitro reconstitution experiments revealed that α-tubulin detyrosination specifically promotes Ndc80 diffusion along the microtubule lattice. We propose that the gradual accumulation of detyrosinated α-tubulin after initial microtubule capture at kinetochores facilitates the biased diffusion of core-binding proteins along the microtubule lattice, establishing a positive feedback loop that sustains kinetochore-microtubule attachments.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_07:00:36.204.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Hugo Osorio
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-21 11:31:57	ID requested
1	2024-10-03 07:06:13	announced
⏵ 2	2024-10-22 07:00:36	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: Kinetochore Microtubule Attachments,Tubulin Detyrosination/Tyrosination code, KMN complex

Helder Maiato
contact affiliation	i3S – Institute for Research and Innovation in Health, University of Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal IBMC – Institute for Molecular and Cell Biology, University of Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal Cell Division Group, Experimental Biology Unit, Department of Biomedicine, Faculty of Medicine, University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto, Portugal
contact email	maiato@i3s.up.pt
lab head
Hugo Osorio
contact affiliation	i3S - Institute for Research and Innovation in Health, University of Porto
contact email	hosorio@i3s.up.pt
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/10/PXD047133

PRIDE project URI

• PRIDE
  1. PXD047133
     1. Label: PRIDE project
     2. Name: Kinetochores read detyrosinated tubulin to sustain microtubule attachments