PXD022703-1

PXD022703 is an original dataset announced via ProteomeXchange.

Title	The deubiquitinase TRABID stabilises the K29/K48-specific E3 ubiquitin ligase HECTD1
Description	The OTU deubiquitinase TRABID is highly tuned for the recognition and cleavage of K29 and K33-linked ubiquitin chains. Yet the cellular functions of these atypical ubiquitin signals remain unclear. Here, we compared the interactome of two different catalytically-dead TRABID constructs, one that lacks the OTU domain and the other a single point mutant that is catalytically inactive. Since both constructs also act as ubiquitin trapping mutants, this approach was used to differentiate between interactors and substrates. The E3 ubiquitin ligase HECTD1 was confirmed as a TRABID substrate, and using UbiCREST and Ubiquitin-AQUA proteomics, we determined that HECTD1 preferentially assembles K29- and K48-linked ubiquitin chains. Further in vitro autoubiquitination assays using ubiquitin mutants established that in contrast to UBE3C, HECTD1 requires the formation of branched K29/K48-linked chains for its full activity. Finally, we used transient knockdown and genetic knock out of TRABID in mammalian cells to show that TRABID stabilises HECTD1 protein levels. This study identifies TRABID-HECTD1 as a K29-specific DUB/E3 pair and provides novel insights on the assembly of branched ubiquitin signals.
HostingRepository	PRIDE
AnnounceDate	2021-06-23
AnnouncementXML	Submission_2021-06-22_22:50:14.638.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Mark Skehel
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidoethyl-L-cysteine; phosphorylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-24 07:52:18	ID requested
⏵ 1	2021-06-22 22:50:15	announced

Dataset with its publication pending

submitter keyword: Ubiquitin, Deubiquitination, Polyubiquitin chain, E3 ubiquitin ligase, ubiquitin thioesterase, Protein degradation, HECTD1, TRABID, K29/K48-linked polyubiquitin chain, lc-ms/ms

Julien D F Licchesi
contact affiliation	Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, UK.
contact email	j.licchesi@bath.ac.uk
lab head
Mark Skehel
contact affiliation	MRC LMB
contact email	mskehel@mrc-lmb.cam.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022703
     1. Label: PRIDE project
     2. Name: The deubiquitinase TRABID stabilises the K29/K48-specific E3 ubiquitin ligase HECTD1