Genome-wide CRISPR screens reveal multitiered mechanisms through which mTORC1 senses mitochondrial dysfunction.

Autor:	Condon KJ; Whitehead Institute for Biomedical Research, Cambridge, MA 02142.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142., Orozco JM; Whitehead Institute for Biomedical Research, Cambridge, MA 02142.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142., Adelmann CH; Whitehead Institute for Biomedical Research, Cambridge, MA 02142.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142., Spinelli JB; Whitehead Institute for Biomedical Research, Cambridge, MA 02142.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142., van der Helm PW; Whitehead Institute for Biomedical Research, Cambridge, MA 02142.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142., Roberts JM; Whitehead Institute for Biomedical Research, Cambridge, MA 02142.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142., Kunchok T; Whitehead Institute for Biomedical Research, Cambridge, MA 02142., Sabatini DM; Whitehead Institute for Biomedical Research, Cambridge, MA 02142; sabatini@wi.mit.edu.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142.; Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, MA 02142.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2021 Jan 26; Vol. 118 (4).
DOI:	10.1073/pnas.2022120118
Abstrakt:	In mammalian cells, nutrients and growth factors signal through an array of upstream proteins to regulate the mTORC1 growth control pathway. Because the full complement of these proteins has not been systematically identified, we developed a FACS-based CRISPR-Cas9 genetic screening strategy to pinpoint genes that regulate mTORC1 activity. Along with almost all known positive components of the mTORC1 pathway, we identified many genes that impact mTORC1 activity, including DCAF7 , CSNK2B , SRSF2 , IRS4 , CCDC43 , and HSD17B10 Using the genome-wide screening data, we generated a focused sublibrary containing single guide RNAs (sgRNAs) targeting hundreds of genes and carried out epistasis screens in cells lacking nutrient- and stress-responsive mTORC1 modulators, including GATOR1, AMPK, GCN2, and ATF4. From these data, we pinpointed mitochondrial function as a particularly important input into mTORC1 signaling. While it is well appreciated that mitochondria signal to mTORC1, the mechanisms are not completely clear. We find that the kinases AMPK and HRI signal, with varying kinetics, mitochondrial distress to mTORC1, and that HRI acts through the ATF4-dependent up-regulation of both Sestrin2 and Redd1. Loss of both AMPK and HRI is sufficient to render mTORC1 signaling largely resistant to mitochondrial dysfunction induced by the ATP synthase inhibitor oligomycin as well as the electron transport chain inhibitors piericidin and antimycin. Taken together, our data reveal a catalog of genes that impact the mTORC1 pathway and clarify the multifaceted ways in which mTORC1 senses mitochondrial dysfunction. Competing Interests: The authors declare no competing interest.
Databáze:	MEDLINE
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