Cell & Bioscience (Jul 2021)

Genome-wide analysis of protein–protein interactions and involvement of viral proteins in SARS-CoV-2 replication

  • Yiling Jiang,
  • Kuijie Tong,
  • Roubin Yao,
  • Yuanze Zhou,
  • Hanwen Lin,
  • Liubing Du,
  • Yunyun Jin,
  • Liu Cao,
  • Jingquan Tan,
  • Xing-Ding Zhang,
  • Deyin Guo,
  • Ji-An Pan,
  • Xiaoxue Peng

DOI
https://doi.org/10.1186/s13578-021-00644-y
Journal volume & issue
Vol. 11, no. 1
pp. 1 – 16

Abstract

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Abstract Background Analysis of viral protein–protein interactions is an essential step to uncover the viral protein functions and the molecular mechanism for the assembly of a viral protein complex. We employed a mammalian two-hybrid system to screen all the viral proteins of SARS-CoV-2 for the protein–protein interactions. Results Our study detected 48 interactions, 14 of which were firstly reported here. Unlike Nsp1 of SARS-CoV, Nsp1 of SARS-CoV-2 has the most interacting partners among all the viral proteins and likely functions as a hub for the viral proteins. Five self-interactions were confirmed, and five interactions, Nsp1/Nsp3.1, Nsp3.1/N, Nsp3.2/Nsp12, Nsp10/Nsp14, and Nsp10/Nsp16, were determined to be positive bidirectionally. Using the replicon reporter system of SARS-CoV-2, we screened all viral Nsps for their impacts on the viral replication and revealed Nsp3.1, the N-terminus of Nsp3, significantly inhibited the replicon reporter gene expression. We found Nsp3 interacted with N through its acidic region at N-terminus, while N interacted with Nsp3 through its NTD, which is rich in the basic amino acids. Furthermore, using purified truncated N and Nsp3 proteins, we determined the direct interactions between Nsp3 and N protein. Conclusions Our findings provided a basis for understanding the functions of coronavirus proteins and supported the potential of interactions as the target for antiviral drug development.

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