An ADAR1 dsRBD3-PKR kinase domain interaction on dsRNA inhibits PKR activation
Adar null mutant mouse embryos die with aberrant double-stranded RNA (dsRNA)-driven interferon induction, and Adar Mavs double mutants, in which interferon induction is prevented, die soon after birth. Protein kinase R (Pkr) is aberrantly activated in Adar Mavs mouse pup intestines before death, intestinal crypt cells die, and intestinal villi are lost. Adar Mavs Eifak2 (Pkr) triple mutant mice rescue all defects and have long-term survival. Adenosine deaminase acting on RNA 1 (ADAR1) and PKR co-immunoprecipitate from cells, suggesting PKR inhibition by direct interaction. AlphaFold studies on an inhibitory PKR dsRNA binding domain (dsRBD)-kinase domain interaction before dsRNA binding and on an inhibitory ADAR1 dsRBD3-PKR kinase domain interaction on dsRNA provide a testable model of the inhibition. Wild-type or editing-inactive human ADAR1 expressed in A549 cells inhibits activation of endogenous PKR. ADAR1 dsRNA binding is required for, but is not sufficient for, PKR inhibition. Mutating the ADAR1 dsRBD3-PKR contact prevents co-immunoprecipitation, ADAR1 inhibition of PKR activity, and co-localization of ADAR1 and PKR in cells.
Authors
- Pavla Linhartová (ORCID: https://orcid.org/0000-0002-9219-357X)
- Ketty Sinigaglia (ORCID: https://orcid.org/0000-0001-5368-6307)
- Radek Malı́k (ORCID: https://orcid.org/0000-0002-6783-1146)
- Lisa Zerad (ORCID: https://orcid.org/0000-0002-6064-7581)
- Anna Cherian (ORCID: https://orcid.org/0000-0003-1072-3440)
- Mary A. O’Connell (ORCID: https://orcid.org/0000-0003-3844-6174)
- Stanislav Stejskal (ORCID: https://orcid.org/0000-0003-0971-343X)
- Liam P. Keegan (ORCID: https://orcid.org/0000-0001-7915-5790)
- Jan Procházka (ORCID: https://orcid.org/0000-0003-4675-8995)
- Dragana Vukić (ORCID: https://orcid.org/0000-0001-9272-728X)
- Valentina Lacovich (ORCID: https://orcid.org/0000-0003-3280-4583)
- Radislav Sedláček (ORCID: https://orcid.org/0000-0002-3352-392X)
- Janka Melicherová
- Nadège Bondurand (ORCID: https://orcid.org/0000-0002-5855-6955)
- Qiupei Du
Institutions
- Central European Institute of Technology (CZ)
- Inserm (FR)
- Université Paris Cité (FR)
- Institute of Molecular Genetics (RU)
- Czech Academy of Sciences, Institute of Molecular Genetics (CZ)
- Institut des Maladies Génétiques Imagine (FR)
Publication Details
- Journal
- Cell Reports
- Published
- 2024-08-01
- DOI
- https://doi.org/10.1016/j.celrep.2024.114618
- Citations
- 29
- Primary Topic
- RNA regulation and disease
- Type
- article
- Field-Weighted Citation Impact
- 6.78