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PXD020667

PXD020667 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleA new role for the DEAD-box RNA helicase DDX3 in co-translational protein quality control
DescriptionDEAD-box RNA helicases are central players in RNA metabolism, however, their role in translation regulation is largely unexplored in parasitic protozoa. Here, we have investigated the role of DDX3 RNA helicase in ribosome-associated protein quality control in Leishmania. We show that ribosomes move more slowly and de novo polypeptide synthesis is reduced in cells lacking DDX3. In accordance with the slowing of ribosome speed, DDX3 depleted cells exhibit higher levels of ribosome-associated ubiquitination. Especially, ubiquitination of nascent polypeptides is enhanced upon DDX3 loss as determined by the isolation of ribosome-associated nascent chains modified either by HA-Ubiquitin or by endogenous ubiquitin using biotinylated-puromycin labeling. Consistent with increased co-translational ubiquitination, quantitative proteomics analysis revealed higher recruitment of E3 ubiquitin ligases and proteasomal components to DDX3 knockout ribosomes to eliminate aberrant nascent polypeptides. In addition, we show that cells lacking DDX3 accumulate cytosolic aggregates. This along with the higher recruitment of ribosome-associated chaperones and the improvement of translation by increasing HSP70 availability suggests that co-translational control of nascent polypeptides is impaired in the absence of DDX3. Altogether, these results highlight an important role for DDX3 in ribosome-associated quality control by reducing co-translational ubiquitination and proteotoxicity, hence allowing optimal ribosome movement and translation elongation.
HostingRepositoryPRIDE
AnnounceDate2021-07-23
AnnouncementXMLSubmission_2021-07-22_23:36:57.271.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterBarbara Papadopoulou
SpeciesList scientific name: Leishmania infantum; NCBI TaxID: 5671;
ModificationListubiquitination signature tetrapeptidyl lysine; ubiquitination signature dipeptidyl lysine
InstrumentOrbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-07-31 01:35:48ID requested
12021-07-22 23:36:57announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: Leishmania infantum, Ribosome, DDX3, Cotranslational protein ubiquitination, Orbitrap Fusion, Label-free quantitative proteomics
Contact List
Barbara Papadopoulou
contact affiliationUniversite Laval-CHU de Quebec Research Center
contact emailbarbara.papadopoulou@crchudequebec.ulaval.ca
lab head
Barbara Papadopoulou
contact affiliationUniversite Laval-CHU de Quebec Research Center
contact emailbarbara.papadopoulou@crchudequebec.ulaval.ca
dataset submitter
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Dataset FTP location
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