Journal
CELL DEATH AND DIFFERENTIATION
Volume 26, Issue 6, Pages 1138-1155Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41418-018-0192-6
Keywords
-
Categories
Funding
- Austrian Academy of Sciences
- Ludwig Boltzmann Society
- European Research Council (ERC AdG) [695214]
- EMBO long-term fellowship [ALTF 1346-2011]
- Marie Sklodowska-Curie fellowship [IEF-301663, IF-661491]
- Austrian Science Fund Lise Meitner Program Fellowship [FWF M1809]
- Austrian Science Fund Grants [FWF F47-B20, FWF I2192, FWF P29250]
- Boehringer Ingelheim [BI-CeMM 238114]
- European Research Council (ERC) [695214] Funding Source: European Research Council (ERC)
- Austrian Science Fund (FWF) [P29250] Funding Source: Austrian Science Fund (FWF)
Ask authors/readers for more resources
Regulation of cell and tissue homeostasis by programmed cell death is a fundamental process with wide physiological and pathological implications. The advent of scalable somatic cell genetic technologies creates the opportunity to functionally map such essential pathways, thereby identifying potential disease-relevant components. We investigated the genetic basis underlying necroptotic cell death by performing a complementary set of loss-of-function and gain-of-function genetic screens. To this end, we established FADD-deficient haploid human KBM7 cells, which specifically and efficiently undergo necroptosis after a single treatment with either TNF alpha or the SMAC mimetic compound birinapant. A series of unbiased gene-trap screens identified key signaling mediators, such as TNFR1, RIPK1, RIPK3, and MLKL. Among the novel components, we focused on the zinc transporter SLC39A7, whose knock-out led to necroptosis resistance by affecting TNF receptor surface levels. Orthogonal, solute carrier (SLC)-focused CRISPR/Cas9-based genetic screens revealed the exquisite specificity of SLC39A7, among similar to 400 SLC genes, for TNFR1-mediated and FAS-mediated but not TRAIL-R1-mediated responses. Mechanistically, we demonstrate that loss of SLC39A7 resulted in augmented ER stress and impaired receptor trafficking, thereby globally affecting downstream signaling. The newly established cellular model also allowed genome-wide gain-of-function screening for genes conferring resistance to necroptosis via the CRISPR/Cas9-based synergistic activation mediator approach. Among these, we found cIAP1 and cIAP2, and characterized the role of TNIP1, which prevented pathway activation in a ubiquitin-binding dependent manner. Altogether, the gain-of-function and loss-of-function screens described here provide a global genetic chart of the molecular factors involved in necroptosis and death receptor signaling, prompting further investigation of their individual contribution and potential role in pathological conditions.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available