Translation stress and collided ribosomes are co-activators of cGAS

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Dokumenter

The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway senses cytosolic DNA and induces interferon-stimulated genes (ISGs) to activate the innate immune system. Here, we report the unexpected discovery that cGAS also senses dysfunctional protein production. Purified ribosomes interact directly with cGAS and stimulate its DNA-dependent activity in vitro. Disruption of the ribosome-associated protein quality control (RQC) pathway, which detects and resolves ribosome collision during translation, results in cGAS-dependent ISG expression and causes re-localization of cGAS from the nucleus to the cytosol. Indeed, cGAS preferentially binds collided ribosomes in vitro, and orthogonal perturbations that result in elevated levels of collided ribosomes and RQC activation cause sub-cellular re-localization of cGAS and ribosome binding in vivo as well. Thus, translation stress potently increases DNA-dependent cGAS activation. These findings have implications for the inflammatory response to viral infection and tumorigenesis, both of which substantially reprogram cellular protein synthesis.

OriginalsprogEngelsk
TidsskriftMolecular Cell
Vol/bind81
Udgave nummer13
Sider (fra-til)2808-2822.e10
ISSN1097-2765
DOI
StatusUdgivet - 2021

Bibliografisk note

Funding Information:
This work was supported by the Francis Crick Institute (FCI receives funding from Cancer Research UK [ FC001166 ], the UK Medical Research Council [ FC001166 ], and the Wellcome Trust [ FC001166 ]), grants to J.Q.S. from the European Research Council (Agreement 693327 ), a Laureate grant from the Novo Nordisk Foundation ( NNF19OC0055875 ), and a Chair grant from the Danish National Research Foundation ( DNRF153 ). Work in the Hegde lab was supported by the UK Medical Research Council ( MC_UP_A022_1007 to R.S.H.). L.W. and Z.H. were supported by the EMBO-LTF program ( EMBO ALTF 1071-2015 and EMBO ALTF 5-2019 , respectively). We thank Anna Herlihy and Jane Walker for assistance with PLA experiments, and members of FCI’s Advanced Sequencing Facility, Cell Services, Flow cytometry facility, and Advanced Light Microscopy for expert technical assistance. The Flp-In compatible destination vectors, Flp-In T-REx U2OS cell line, and STING cDNA were kindly provided by Markus Landthaler, Erich Nigg, and Pinglong Xu, respectively. We thank Qian Zhang, Annemarthe Van der Veen, Joan Manils, Subramanian Venkatesan, Marie-Luise Winz, Yisui Xia, Zhewang Lin, Shuyang Chen, and Viswanathan Chandrasekaran for their input and all Svejstrup lab members for discussions. We thank Barbara Dirac-Svejstrup, Peter Verrijzer, and David Lopez Martinez for comments on the manuscript.

Funding Information:
This work was supported by the Francis Crick Institute (FCI receives funding from Cancer Research UK [FC001166], the UK Medical Research Council [FC001166], and the Wellcome Trust [FC001166]), grants to J.Q.S. from the European Research Council (Agreement 693327), a Laureate grant from the Novo Nordisk Foundation (NNF19OC0055875), and a Chair grant from the Danish National Research Foundation (DNRF153). Work in the Hegde lab was supported by the UK Medical Research Council (MC_UP_A022_1007 to R.S.H.). L.W. and Z.H. were supported by the EMBO-LTF program (EMBO ALTF 1071-2015 and EMBO ALTF 5-2019, respectively). We thank Anna Herlihy and Jane Walker for assistance with PLA experiments, and members of FCI's Advanced Sequencing Facility, Cell Services, Flow cytometry facility, and Advanced Light Microscopy for expert technical assistance. The Flp-In compatible destination vectors, Flp-In T-REx U2OS cell line, and STING cDNA were kindly provided by Markus Landthaler, Erich Nigg, and Pinglong Xu, respectively. We thank Qian Zhang, Annemarthe Van der Veen, Joan Manils, Subramanian Venkatesan, Marie-Luise Winz, Yisui Xia, Zhewang Lin, Shuyang Chen, and Viswanathan Chandrasekaran for their input and all Svejstrup lab members for discussions. We thank Barbara Dirac-Svejstrup, Peter Verrijzer, and David Lopez Martinez for comments on the manuscript. L.W. and J.Q.S. conceived the project. L.W. performed the majority of experiments, but S.J. P.K.B. Z.H. and P.F. helped L.W. generate collided ribosomes, purify recombinant cGAS, establish the cGAM in vitro assay, and perform mass spectrometry on interactomes, respectively. D.B. performed and analyzed flow cytometry experiments and also performed in vivo cGAMP measurements. R.M. analyzed TTchem-seq data. A.S. A.P.S. R.S.H. and J.Q.S. supervised the work. L.W. and J.Q.S. wrote the manuscript, with input from all authors. The authors declare no competing interests.

Publisher Copyright:
© 2021 The Author(s)

Antal downloads er baseret på statistik fra Google Scholar og www.ku.dk


Ingen data tilgængelig

ID: 275822778