title: Cooperative assembly of IFI16 filaments on dsDNA provides insights into host defense strategy.
abstract: Whether host DNA receptors have any capacity to distinguish self from nonself at the molecular level is an outstanding question in the innate immunity of mammals. Here, by using quantitative assays and electron microscopy, we show that cooperatively assembling into filaments on dsDNA may serve as an integral mechanism by which human IFN-inducible protein-16 (IFI16) engages foreign DNA. IFI16 is essential for defense against a number of different pathogens, and its aberrant activity is also implicated in several autoimmune disorders, such as Sjögren syndrome. IFI16 cooperatively binds dsDNA in a length-dependent manner and clusters into distinct protein filaments even in the presence of excess dsDNA. Consequently, the assembled IFI16⋅dsDNA oligomers are clearly different from the conventional noninteracting entities resembling beads on a string. The isolated DNA-binding domains of IFI16 engage dsDNA without forming filaments and with weak affinity, and it is the non-DNA-binding pyrin domain of IFI16 that drives the cooperative filament assembly. The surface residues on the pyrin domain that mediate the cooperative DNA binding are conserved, suggesting that related receptors use a common mechanism. These results suggest that IFI16 clusters into signaling foci in a switch-like manner and that it is capable of using the size of naked dsDNA as a molecular ruler to distinguish self from nonself.
authors.affiliation: Departments of Biophysics and Biophysical Chemistry and Cell Biology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205.
authors.forename: Seamus R
authors.initials: SR
authors.lastname: Morrone
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.forename: Leeza M
authors.initials: LM
authors.lastname: Constantoulakis
authors.forename: Richard M
authors.initials: RM
authors.lastname: Hooy
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Delannoy
authors.forename: Jungsan
authors.initials: J
authors.lastname: Sohn
mesh_terms: D000595:Amino Acid Sequence; D001667:Binding, Competitive; D002467:Cell Nucleus / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D006801:Humans; D007113:Immunity, Innate; D007249:Inflammation; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015398:Signal Transduction
chemical_list: D003432:Cross-Linking Reagents; D009687:Nuclear Proteins; D010750:Phosphoproteins; C076700:IFI16 protein, human; D004247:DNA
keywords: cooperative filament formation; inflammasome