CRISPR screens identify novel regulators of cFLIP dependency and ligand-independent, TRAIL-R1-mediated cell death.
Journal
Cell death and differentiation
ISSN: 1476-5403
Titre abrégé: Cell Death Differ
Pays: England
ID NLM: 9437445
Informations de publication
Date de publication:
05 2023
05 2023
Historique:
received:
16
08
2022
accepted:
10
02
2023
revised:
07
02
2023
pmc-release:
01
05
2024
medline:
4
5
2023
pubmed:
22
2
2023
entrez:
21
2
2023
Statut:
ppublish
Résumé
Kaposi's sarcoma-associated herpesvirus (KSHV) causes primary effusion lymphoma (PEL). PEL cell lines require expression of the cellular FLICE inhibitory protein (cFLIP) for survival, although KSHV encodes a viral homolog of this protein (vFLIP). Cellular and viral FLIP proteins have several functions, including, most importantly, the inhibition of pro-apoptotic caspase 8 and modulation of NF-κB signaling. To investigate the essential role of cFLIP and its potential redundancy with vFLIP in PEL cells, we first performed rescue experiments with human or viral FLIP proteins known to affect FLIP target pathways differently. The long and short isoforms of cFLIP and molluscum contagiosum virus MC159L, which are all strong caspase 8 inhibitors, efficiently rescued the loss of endogenous cFLIP activity in PEL cells. KSHV vFLIP was unable to fully rescue the loss of endogenous cFLIP and is therefore functionally distinct. Next, we employed genome-wide CRISPR/Cas9 synthetic rescue screens to identify loss of function perturbations that can compensate for cFLIP knockout. Results from these screens and our validation experiments implicate the canonical cFLIP target caspase 8 and TRAIL receptor 1 (TRAIL-R1 or TNFRSF10A) in promoting constitutive death signaling in PEL cells. However, this process was independent of TRAIL receptor 2 or TRAIL, the latter of which is not detectable in PEL cell cultures. The requirement for cFLIP is also overcome by inactivation of the ER/Golgi resident chondroitin sulfate proteoglycan synthesis and UFMylation pathways, Jagunal homolog 1 (JAGN1) or CXCR4. UFMylation and JAGN1, but not chondroitin sulfate proteoglycan synthesis or CXCR4, contribute to TRAIL-R1 expression. In sum, our work shows that cFLIP is required in PEL cells to inhibit ligand-independent TRAIL-R1 cell death signaling downstream of a complex set of ER/Golgi-associated processes that have not previously been implicated in cFLIP or TRAIL-R1 function.
Identifiants
pubmed: 36801923
doi: 10.1038/s41418-023-01133-0
pii: 10.1038/s41418-023-01133-0
pmc: PMC10154404
doi:
Substances chimiques
CASP8 and FADD-Like Apoptosis Regulating Protein
0
Caspase 8
EC 3.4.22.-
JAGN1 protein, human
0
Ligands
0
Membrane Proteins
0
Proteoglycans
0
Sulfates
0
TNFRSF11A protein, human
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
1221-1234Subventions
Organisme : NCI NIH HHS
ID : K22 CA241355
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA247619
Pays : United States
Organisme : NIAID NIH HHS
ID : T32 AI007476
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA060553
Pays : United States
Informations de copyright
© 2023. The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare.
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