title: Crystal structure of human ISG15 protein in complex with influenza B virus NS1B.
abstract: ISG15 (interferon-stimulated gene 15), the first ubiquitin-like protein (UBL) identified, has emerged as an important cellular antiviral factor. It consists of two UBL domains with a short linker between them. The covalent attachment of ISG15 to host and viral proteins to modify their functions, similar to ubiquitylation, is named ISGylation. Influenza B virus NS1B protein antagonizes human but not mouse ISGylation because NS1B exhibits species specificity; it only binds human and non-human primate ISG15. Previous studies have demonstrated that the N-terminal UBL domain and linker of ISG15 are required for the binding by NS1B and that the linker plays a large role in the species specificity, but the structural basis for them has not been elucidated. Here we report the crystal structure of human ISG15 in complex with NS1B at a resolution of 2.0 Å. A loop in the ISG15 N-terminal UBL domain inserts into a pocket in the NS1B dimer, forming a high affinity binding site. The nonspecific van der Waals contacts around the ISG15 linker form a low affinity site for NS1B binding. However, sequence alignment reveals that residues in the high affinity site are highly conserved in primate and non-primate ISG15. We propose that the low affinity binding around the ISG15 linker is important for the initial contact with NS1B and that the stable complex formation is largely contributed by the following high affinity interactions between ISG15 N-terminal UBL domain and NS1B. This provides a structural basis for the species-specific binding of ISG15 by the NS1B protein.
authors.affiliation: Center for Structural Biology, Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084 and.
authors.forename: Liang
authors.initials: L
authors.lastname: Li
authors.affiliation: Center for Structural Biology, Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084 and.
authors.forename: Dongli
authors.initials: D
authors.lastname: Wang
authors.affiliation: Center for Structural Biology, Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084 and.
authors.forename: Yinan
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Center for Structural Biology, Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084 and.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Sun
authors.affiliation: Center for Structural Biology, Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084 and.
authors.forename: Senyan
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Center for Structural Biology, Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084 and. Electronic address: xinquanwang@mail.tsinghua.edu.cn.
authors.forename: Xinquan
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D001665:Binding Sites; D002850:Chromatography, Gel; D018360:Crystallography, X-Ray / Q000379:methods; D016207:Cytokines / Q000737:chemistry*; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D009981:Influenza B virus / Q000378:metabolism*; D051379:Mice; D008968:Molecular Conformation; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D014452:Ubiquitins / Q000737:chemistry*; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D016207:Cytokines; C085820:INS1 protein, influenza virus; D011994:Recombinant Proteins; D014452:Ubiquitins; D017361:Viral Nonstructural Proteins; C074034:ISG15 protein, human