Structural basis for IFN antagonism by human respiratory syncytial virus nonstructural protein 2. 2021

Jingjing Pei, and Nicole D Wagner, and Angela J Zou, and Srirupa Chatterjee, and Dominika Borek, and Aidan R Cole, and Preston J Kim, and Christopher F Basler, and Zbyszek Otwinowski, and Michael L Gross, and Gaya K Amarasinghe, and Daisy W Leung
John T. Milliken Department of Medicine, Division of Infectious Diseases, Washington University School of Medicine, St. Louis, MO 63110.

Human respiratory syncytial virus (RSV) nonstructural protein 2 (NS2) inhibits host interferon (IFN) responses stimulated by RSV infection by targeting early steps in the IFN-signaling pathway. But the molecular mechanisms related to how NS2 regulates these processes remain incompletely understood. To address this gap, here we solved the X-ray crystal structure of NS2. This structure revealed a unique fold that is distinct from other known viral IFN antagonists, including RSV NS1. We also show that NS2 directly interacts with an inactive conformation of the RIG-I-like receptors (RLRs) RIG-I and MDA5. NS2 binding prevents RLR ubiquitination, a process critical for prolonged activation of downstream signaling. Structural analysis, including by hydrogen-deuterium exchange coupled to mass spectrometry, revealed that the N terminus of NS2 is essential for binding to the RIG-I caspase activation and recruitment domains. N-terminal mutations significantly diminish RIG-I interactions and result in increased IFNβ messenger RNA levels. Collectively, our studies uncover a previously unappreciated regulatory mechanism by which NS2 further modulates host responses and define an approach for targeting host responses.

UI MeSH Term Description Entries
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D011971 Receptors, Immunologic Cell surface molecules on cells of the immune system that specifically bind surface molecules or messenger molecules and trigger changes in the behavior of cells. Although these receptors were first identified in the immune system, many have important functions elsewhere. Immunologic Receptors,Immunologic Receptor,Immunological Receptors,Receptor, Immunologic,Receptors, Immunological
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000071457 DEAD Box Protein 58 A DEAD-box RNA helicase that contains an N-terminal DEATH-LIKE DOMAIN, AAA+ ATPase domain, and C-terminal RNA HELICASE activity. It functions as an innate immune receptor through its recognition of viral nucleic acids. It also induces the expression of INTERFERON TYPE I and proinflammatory CYTOKINES. Its ligands include: 5'-triphosphorylated SINGLE-STRANDED RNA, DOUBLE-STRANDED RNA (dsRNA), and short dsRNA (less than 1 kb in length). DEAD (Asp-Glu-Ala-Asp) Box Polypeptide 58,Probable ATP-Dependent RNA Helicase DDX58,RIG-I-like Receptor 1,Probable ATP Dependent RNA Helicase DDX58,RIG I like Receptor 1
D000072640 Interferon-Induced Helicase, IFIH1 A DEAD box RNA helicase that contains two N-terminal CASPASE ACTIVATION AND RECRUITMENT DOMAINS. It functions as a sensor of viral NUCLEIC ACIDS such as DOUBLE-STRANDED RNA and activates the INNATE IMMUNE RESPONSE by inducing the expression of INTERFERON-ALPHA and INTERFERON-BETA. It may also regulate cell growth and APOPTOSIS. CADM-140 Autoantigen,Helicard,IFIH1 Protein,Interferon Induced with Helicase C Domain 1,Interferon-Induced Helicase C Domain-Containing Protein 1,Melanoma Differentiation-Associated Protein 5,Autoantigen, CADM-140,CADM 140 Autoantigen,Helicase, IFIH1 Interferon-Induced,IFIH1 Interferon-Induced Helicase,Interferon Induced Helicase C Domain Containing Protein 1,Interferon Induced Helicase, IFIH1,Melanoma Differentiation Associated Protein 5
D012333 RNA, Messenger RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm. Messenger RNA,Messenger RNA, Polyadenylated,Poly(A) Tail,Poly(A)+ RNA,Poly(A)+ mRNA,RNA, Messenger, Polyadenylated,RNA, Polyadenylated,mRNA,mRNA, Non-Polyadenylated,mRNA, Polyadenylated,Non-Polyadenylated mRNA,Poly(A) RNA,Polyadenylated mRNA,Non Polyadenylated mRNA,Polyadenylated Messenger RNA,Polyadenylated RNA,RNA, Polyadenylated Messenger,mRNA, Non Polyadenylated
D016899 Interferon-beta One of the type I interferons produced by fibroblasts in response to stimulation by live or inactivated virus or by double-stranded RNA. It is a cytokine with antiviral, antiproliferative, and immunomodulating activity. Interferon, Fibroblast,beta-Interferon,Fiblaferon,Interferon, beta,Interferon, beta-1,Interferon-beta1,beta-1 Interferon,Fibroblast Interferon,Interferon beta,Interferon beta1,Interferon, beta 1,beta 1 Interferon,beta Interferon
D017361 Viral Nonstructural Proteins Proteins encoded by a VIRAL GENOME that are not structural components of VIRUS PARTICLES. Some of these proteins may play roles within the infected cell during VIRUS REPLICATION or act in regulation of virus replication or VIRUS ASSEMBLY. Nonstructural Proteins, Viral,NS Proteins, Viral,Viral NS Proteins,Viral Non-Structural Proteins,Viral Nonstructural Protein,Viral Nonstructural Proteins NS1,Viral Nonstructural Proteins NS2,Nonstructural Protein, Viral,Viral Non Structural Proteins
D041961 Deuterium Exchange Measurement A research technique to measure solvent exposed regions of molecules that is used to provide insight about PROTEIN CONFORMATION. Hydrogen-Deuterium Exchange Measurement,Deuterium Exchange Measurements,Exchange Measurement, Deuterium,Exchange Measurement, Hydrogen-Deuterium,Exchange Measurements, Deuterium,Exchange Measurements, Hydrogen-Deuterium,Hydrogen Deuterium Exchange Measurement,Hydrogen-Deuterium Exchange Measurements,Measurement, Deuterium Exchange,Measurement, Hydrogen-Deuterium Exchange,Measurements, Deuterium Exchange,Measurements, Hydrogen-Deuterium Exchange
D057809 HEK293 Cells A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5. 293T Cells,HEK 293 Cell Line,HEK 293 Cells,Human Embryonic Kidney Cell Line 293,Human Kidney Cell Line 293,293 Cell, HEK,293 Cells, HEK,293T Cell,Cell, 293T,Cell, HEK 293,Cell, HEK293,Cells, 293T,Cells, HEK 293,Cells, HEK293,HEK 293 Cell,HEK293 Cell

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