PXD043646

PXD043646 is an original dataset announced via ProteomeXchange.

Title	Cryo-EM visualizes substrate channeling and E2 multisite phosphorylation configuring ubiquitin ligation by GID/CTLH E3
Description	Ubiquitylation is catalyzed by coordinated actions of E3 and E2 enzymes. Molecular principles governing many important E3-E2 partnerships remain unknown, including for RING-family GID/CTLH E3 ubiquitin ligases and their dedicated E2, Ubc8/UBE2H (yeast/human nomenclature). GID/CTLH-Ubc8/UBE2H-mediated ubiquitylation regulates biological processes ranging from yeast metabolic signaling to human development. Here, cryo-EM, biochemistry, and cell biology reveal exquisitely specific GID/CTLH-Ubc8/UBE2H pairing through an unconventional catalytic assembly and auxiliary interactions 70-100 Å away, mediated by E2 phosphorylation. Rather than dynamic polyelectrostatic interactions reported for other ubiquitylation complexes, multiple Ubc8/UBE2H phosphorylation sites within acidic CK2-targeted sequences specifically anchor the E2 C-termini to E3 basic patches. Positions of phospho-dependent interactions relative to the catalytic domains correlate across evolution. Overall, our data illustrate phosphorylation-dependent multivalency establishes a specific E3-E2 partnership, is antagonistic with dephosphorylation, immobilizes the catalytic centers within a flexing GID E3-substrate assembly, and facilitates substrate collision with ubiquitylation active sites.
HostingRepository	PRIDE
AnnounceDate	2023-12-18
AnnouncementXML	Submission_2023-12-18_09:03:44.807.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Mario Oroshi
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-07-10 08:47:45	ID requested
⏵ 1	2023-12-18 09:03:45	announced

Dataset with its publication pending

submitter keyword: Proteomics

Matthias Mann
contact affiliation	Research Department "Proteomics and Signal Transduction"
contact email	mmann@biochem.mpg.de
lab head
Mario Oroshi
contact affiliation	Proteomics
contact email	oroshi@biochem.mpg.de
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/2023/12/PXD043646

PRIDE project URI

• PRIDE
  1. PXD043646
     1. Label: PRIDE project
     2. Name: Cryo-EM visualizes substrate channeling and E2 multisite phosphorylation configuring ubiquitin ligation by GID/CTLH E3