Structural basis for cytosolic double-stranded RNA surveillance by human oligoadenylate synthetase 1. Author Jesse Donovan, Matthew Dufner, Alexei Korennykh Publication Year 2013 Type Journal Article Abstract The human sensor of double-stranded RNA (dsRNA) oligoadenylate synthetase 1 (hOAS1) polymerizes ATP into 2',5'-linked iso-RNA (2-5A) involved in innate immunity, cell cycle, and differentiation. We report the crystal structure of hOAS1 in complex with dsRNA and 2'-deoxy ATP at 2.7 Å resolution, which reveals the mechanism of cytoplasmic dsRNA recognition and activation of oligoadenylate synthetases. Human OAS1 recognizes dsRNA using a previously uncharacterized protein/RNA interface that forms via a conformational change induced by binding of dsRNA. The protein/RNA interface involves two minor grooves and has no sequence-specific contacts, with the exception of a single hydrogen bond between the -NH(2) group of nucleobase G17 and the carbonyl oxygen of serine 56. Using a biochemical readout, we show that hOAS1 undergoes more than 20,000-fold activation upon dsRNA binding and that canonical or GU-wobble substitutions produce dsRNA mutants that retain either full or partial activity, in agreement with the crystal structure. Ultimately, the binding of dsRNA promotes an elaborate conformational rearrangement in the N-terminal lobe of hOAS1, which brings residues D75, D77, and D148 into proximity and creates coordination geometry for binding of two catalytic Mg(2+) ions and ATP. The assembly of this critical active-site structure provides the gate that couples binding of dsRNA to the production and downstream functions of 2-5A. Keywords Base Sequence, Molecular Sequence Data, Nucleic Acid Conformation, Humans, Binding Sites, Substrate Specificity, Protein Binding, Models, Molecular, Protein Structure, Tertiary, Crystallography, X-Ray, Amino Acid Sequence, Sequence Homology, Amino Acid, Enzyme Activation, Biocatalysis, 2',5'-Oligoadenylate Synthetase, Adenine Nucleotides, Oligoribonucleotides, RNA, Double-Stranded, Cytosol, Deoxyadenine Nucleotides Journal Proc Natl Acad Sci U S A Volume 110 Issue 5 Pages 1652-7 Date Published 2013 Jan 29 ISSN Number 1091-6490 DOI 10.1073/pnas.1218528110 Alternate Journal Proc Natl Acad Sci U S A PMCID PMC3562804 PMID 23319625 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML