PXD043060

PXD043060 is an original dataset announced via ProteomeXchange.

Title	PTP1B inhibition promotes cell death in hypoxia by triggering RNF213-dependent pyroptosis
Description	The tumor microenvironment (TME) typically includes hypoxic regions. Hypoxic cancer cells are resistant to many anti-neoplastic therapies and can seed recurrence; consequently, understanding how they survive could yield new therapeutic strategies. In response to decreasing oxygen, cells are known to successively activate hypoxia-inducible factor 1-alpha (HIF1alpha), endoplasmic reticulum (ER) stress, and AMP-dependent protein kinase (AMPK). We previously identified an additional pathway in which the protein-tyrosine phosphatase PTP1B (encoded by PTPN1) prevents the death of HER2+ breast cancer cells exposed to severe hypoxia (O2<1%) by inhibiting RNF213, a large (~600 kDa) E3 ubiquitin ligase containing two functional AAA-ATPase domains and two ubiquitin ligase domains (RING and RZ) that also is implicated in Moyamoya disease (MMD), lipotoxicity, and innate immunity. These findings provided a potential explanation for earlier observations that Ptpn1-/- mice are resistant to Neu (rodent HER2)-induced breast cancer, but how PTP1B regulates RNF213, the identity(ies) of RNF213 targets, the mechanism of cell death, and the relevance to other RNF213 actions remained unclear. Via proximity-based proteomics, we identified the CYLD-SPATA2 complex, a major negative regulator of NF-kappaB activation, as an RNF213 substrate critical for hypoxia-induced cell death. Here we show that PTP1B and ABL1/2 reciprocally control the phosphorylation of RNF213 on Tyrosine-1275. Phosphorylation of this site promotes RNF213 oligomerization, resulting in RZ domain activation, LUBAC complex-dependent ubiquitylation and degradation of the CYLD/SPATA2 deubiquitinase complex, and NF-kappaB activation. NF-kappaB induces NLRP3 inflammasome priming, and concomitant hypoxia-induced ER stress triggers inflammasome activation, resulting in pyroptotic cell death. Mutagenesis experiments show that the RNF213 RING negatively regulates the RZ domain, and RING domain mutants, including those associated with MMD, catalyze LUBAC-independent CYLD/SPATA2 degradation. Our results identify a novel PTP1B/RNF213/CYLD/SPATA2 pathway responsible for the pyroptotic death of hypoxic tumor cells, reveal new insights into RNF213 regulation, and have potentially important implications for the pathogenesis of MMD, atherosclerosis, and inflammatory and auto-immune disorders. The mass spectrometry raw files for the phosphorylation identification and proximity labeling results are included here.
HostingRepository	MassIVE
AnnounceDate	2024-08-12
AnnouncementXML	Submission_2024-08-12_08:59:59.334.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Beatrix Ueberheide
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationList	Phospho; Oxidation
Instrument	Q Exactive; Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2023-06-16 08:27:13	ID requested
⏵ 1	2024-08-12 08:59:59	announced

no publication

submitter keyword: RNF213, Proximity Labeling, Turbo ID, phosphorylation

Beatrix Ueberheide
contact affiliation	NYU School of Medicine
contact email	beatrix.ueberheide@nyulangone.org
lab head
Beatrix Ueberheide
contact affiliation	NYU School of Medicine
contact email	beatrixueberheide@gmail.com
dataset submitter

MassIVE dataset URI

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