PXD035503

PXD035503 is an original dataset announced via ProteomeXchange.

Title	The E3 ubiquitin ligase HECTD1 contributes to cell proliferation through an effect on mitosis
Description	Mechanistic studies of how protein ubiquitylation regulates the cell cycle, in particular during mitosis, have provided unique insights which have contributed to the emergence of the ‘Ubiquitin code’. In contrast to RING E3 ubiquitin ligases such as the APC/c ligase complex, the contribution of other E3 ligase families during cell cycle progression remains less well understood. Similarly, the roles of ubiquitin chain types beyond homotypic K48 chains in S-phase or branched K11/K48 chains during mitosis, also remains to be fully determined. Our recent findings that HECTD1 ubiquitin ligase activity assembles branched K29/K48 ubiquitin linkages prompted us to evaluate its function during the cell cycle. We used transient knockdown and genetic knockout to show that HECTD1 depletion in HEK293T and HeLa cells decreases cell number and we established that this is mediated through loss of ubiquitin ligase activity. Interestingly, we found that HECTD1 depletion increases the proportion of cells with aligned chromosomes (Prometa/Metaphase) and we confirmed this molecularly using phospho-Histone H3 (Ser28) as a marker of mitosis. Time-lapse microscopy of NEBD to anaphase onset established that HECTD1-depleted cells take on average longer to go through mitosis. To explore the underlying mechanisms, we used proteomics and identified the interactome of endogenous HECTD1 in cells synchronized in mitosis and validated the Mitosis Checkpoint Complex protein BUB3 as a novel HECTD1 interactor. In line with this, we found that HECTD1 depletion reduces the activity of the Spindle Assembly Checkpoint. Overall, our data suggests a novel role for HECTD1 ubiquitin ligase activity during mitosis.
HostingRepository	PRIDE
AnnounceDate	2022-08-03
AnnouncementXML	Submission_2022-08-03_09:00:07.940.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Holger Kramer
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-21 08:19:59	ID requested
⏵ 1	2022-08-03 09:00:08	announced

Vaughan N, Scholz N, Lindon C, Licchesi JDF, The E3 ubiquitin ligase HECTD1 contributes to cell proliferation through an effect on mitosis. Sci Rep, 12(1):13160(2022) [pubmed]

submitter keyword: Ubiquitin, HECTD1, E3 ubiquitin ligase, cell cycle, cell proliferation, TRABID

Mark Skehel
contact affiliation	The Francis Crick Institute 1 Midland Road London, NW1 1AT United Kingdom
contact email	mark.skehel@crick.ac.uk
lab head
Holger Kramer
contact affiliation	MRC London Institute of Medical Sciences
contact email	h.kramer@lms.mrc.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD035503
     1. Label: PRIDE project
     2. Name: The E3 ubiquitin ligase HECTD1 contributes to cell proliferation through an effect on mitosis