6IVD

TGEV nsp1 mutant - 91-95sg

6IVD の概要

• エントリーDOI: 10.2210/pdb6ivd/pdb
• 分子名称: nsp1 mutant protein (2 entities in total)
• 機能のキーワード: transmissible gastroenteritis virus, nsp1 mutant, viral protein
• 由来する生物種: Transmissible gastroenteritis virus
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 24363.88

構造登録者

Shen, Z.,Peng, G.Q. (登録日: 2018-12-03, 公開日: 2019-08-07, 最終更新日: 2024-03-27)

主引用文献

Shen, Z.,Wang, G.,Yang, Y.,Shi, J.,Fang, L.,Li, F.,Xiao, S.,Fu, Z.F.,Peng, G.
A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence.
J.Biol.Chem., 294:13606-13618, 2019

PubMed Abstract: Coronaviruses are enveloped, single-stranded RNA viruses that are distributed worldwide. They include transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), and the human coronaviruses severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV), many of which seriously endanger human health and well-being. Only alphacoronaviruses and betacoronaviruses harbor nonstructural protein 1 (nsp1), which performs multiple functions in inhibiting antiviral host responses. The role of the C terminus of betacoronavirus nsp1 in virulence has been characterized, but the location of the alphacoronavirus nsp1 region that is important for virulence remains unclear. Here, using TGEV nsp1 as a model to explore the function of this protein in alphacoronaviruses, we demonstrate that alphacoronavirus nsp1 inhibits host gene expression. Solving the crystal structure of full-length TGEV at 1.85-Å resolution and conducting several biochemical analyses, we observed that a specific motif (amino acids 91-95) of alphacoronavirus nsp1 is a conserved region that inhibits host protein synthesis. Using a reverse-genetics system based on CRISPR/Cas9 technology to construct a recombinant TGEV in which this specific nsp1 motif was altered, we found that this mutation does not affect virus replication in cell culture but significantly reduces TGEV pathogenicity in pigs. Taken together, our findings suggest that alphacoronavirus nsp1 is an essential virulence determinant, providing a potential paradigm for the development of a new attenuated vaccine based on modified nsp1.

PubMed: 31350335
DOI: 10.1074/jbc.RA119.009713

実験手法

X-RAY DIFFRACTION (1.975 Å)