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PXD044816

PXD044816 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitlePML::RARA and GATA2 proteins interact via DNA templates to induce aberrant self-renewal in hematopoietic cells
DescriptionThe global protein interactions of the oncofusion protein PML::RARA were identified using proximity labeling followed by mass spectrometry. Briefly, Lineage-depleted bone marrow cells from C57Bl/6 mice were transduced with MSCV-IRES-GFP retroviruses containing an enhanced biotin ligase (TurboID) fused to either the N-terminus or C-terminus of PML::RARAWT (TurboID-PML::RARAWT and PML::RARAWT-TurboID respectively), TurboID-PML::RARAC88A (mutation in the DNA binding domain of RARA), or a TurboID cDNA alone. A flexible Glycine/Serine linker placed between TurboID and PML::RARA allows TurboID to freely rotate, and biotinylate proteins within 10 angstroms of the TurboID-PML::RARA fusion protein. Four days following the first round of transduction, cells were flow sorted on GFP+ cells. Two days following the sort, 1-5 million cells were treated with 50 uM Biotin for four hours at 37 degrees C. Cell lysates were prepared, sonicated, and incubated with streptavidin agarose beads overnight. Beads were washed, peptides were eluted and digested, and then run on a timsTOF Pro 2.
HostingRepositoryMassIVE
AnnounceDate2024-04-05
AnnouncementXMLSubmission_2024-04-05_11:29:51.993.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterJim Malone
SpeciesList scientific name: Mus musculus; common name: house mouse; NCBI TaxID: 10090;
ModificationListCarbamidomethyl; Xlink:DTSSP[88]; Deamidated; Gln->pyro-Glu; Oxidation
InstrumenttimsTOF Pro
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-08-23 20:20:45ID requested
12024-04-05 11:29:52announced
Publication List
no publication
Keyword List
submitter keyword: PML::RARA, TurboID, Proximity labeling, Biotin ligase, mass spectrometry
Contact List
Timothy J. Ley
contact affiliationWashington University in St. Louis
contact emailtimley@wustl.edu
lab head
Jim Malone
contact affiliationWashington University Sch
contact emailjmalone@wustl.edu
dataset submitter
Full Dataset Link List
MassIVE dataset URI
Dataset FTP location
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