5HIY

Crystal structure of PEDV NSP9 Mutant-C59A

5HIY の概要

• エントリーDOI: 10.2210/pdb5hiy/pdb
• 関連するPDBエントリー: 5HIZ
• 分子名称: Non-structural protein 9 (1 entity in total)
• 機能のキーワード: rna binding proteins, rna binding protein
• 由来する生物種: Porcine epidemic diarrhea virus CV777 (PEDV)
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 38714.01

構造登録者

Deng, F.,Peng, G. (登録日: 2016-01-12, 公開日: 2017-01-25, 最終更新日: 2024-03-20)

主引用文献

Zeng, Z.,Deng, F.,Shi, K.,Ye, G.,Wang, G.,Fang, L.,Xiao, S.,Fu, Z.,Peng, G.
Dimerization of Coronavirus nsp9 with Diverse Modes Enhances Its Nucleic Acid Binding Affinity.
J.Virol., 92:-, 2018

PubMed Abstract: Coronaviruses pose serious health threats to humans and other animals. Understanding the mechanisms of their replication has important implications for global health and economic stability. Nonstructural protein 9 (nsp9) is an essential RNA binding protein for coronavirus replication. However, the mechanisms of the dimerization and nucleic acid binding of nsp9 remain elusive. Here, we report four crystal structures, including wild-type porcine delta coronavirus (PDCoV) nsp9, PDCoV nsp9-ΔN7 (N-terminal 7 amino acids deleted), wild-type porcine epidemic diarrhea virus (PEDV) nsp9, and PEDV nsp9-C59A mutant. These structures reveal the diverse dimerization forms of coronavirus nsp9. We first found that the N-finger of nsp9 from PDCoV plays a critical role in dimerization. Meanwhile, PEDV nsp9 is distinguished by the presence of a disulfide bond in the dimer interface. Interestingly, size exclusion chromatography and analytical ultracentrifugation analyses indicate that the PDCoV nsp9-ΔN7 and PEDV nsp9-C59A mutants are monomeric in solution. In addition, electrophoretic mobility shift assays and microscale thermophoresis analysis indicate that the monomeric forms of PDCoV nsp9 and PEDV nsp9 still have nucleic acid binding affinity, although it is lower than that of the wild type. Our results show that the diverse dimerization forms of coronavirus nsp9 proteins enhance their nucleic acid binding affinity. Coronaviruses cause widespread respiratory, gastrointestinal, and central nervous system diseases in humans and other animals, threatening human health and causing economic loss. Coronavirus nsp9, a member of the replication complex, is an important RNA binding subunit in the RNA-synthesizing machinery of all coronaviruses. However, the mechanisms of the dimerization and nucleic acid binding of nsp9 remain elusive. In this study we determined the nsp9 crystal structures of PDCoV and PEDV. We first found that the N-finger of nsp9 from PDCoV plays a critical role in dimerization. Meanwhile, PEDV nsp9 is distinguished by the presence of a disulfide bond in the dimer interface. This study provides a structural and functional basis for understanding the mechanism of dimerization and shows that the diverse dimerization modes of coronavirus nsp9 proteins enhance their nucleic acid binding affinity. Importantly, these findings may provide a new insight for antiviral drug development.

PubMed: 29925659
DOI: 10.1128/JVI.00692-18

実験手法

X-RAY DIFFRACTION (3 Å)