Generation of recombinant influenza virus bearing strep tagged PB2 and effective identification of interactional host factors

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Author(s)

  • Huanan Li
  • Yuandi Yu
  • Xingxing Ren
  • Bo Li
  • Jiahao Zhang
  • Ming Liao
  • Wenbao Qi

Related Research Unit(s)

Detail(s)

Original languageEnglish
Article number108985
Journal / PublicationVeterinary Microbiology
Volume254
Online published13 Jan 2021
Publication statusPublished - Mar 2021

Abstract

The genome of influenza A virus is negative-sense and segmented RNA, which is transcribed and replicated by the viral RNA-dependent RNA polymerase (RdRp) during the virus life cycle. The viral RdRp is thought to be an important host range and virulence determinant factor, and the 627 site of PB2 subunit is a highly acceptable key site of RdRp function. Besides, the function of RdRp is modulated by several host factors. Identification of the host factors interacting with RdRp is of great interest. Here, we tried to explore an effective method to study virus-host interaction by rescuing replication-competent recombinant influenza viruses carrying Strep tagged PB2. Subsequently, we tested several biological characteristics of recombinant viruses in cells and pathogenicity in mice. Then, we purified of protein complex of Strep tagged PB2 and host factors of interest from 293 T cells infected with recombinant viruses. After purification, we performed mass spectrometry to identify these proteins that interacting with PB2. We identified 57 host factors in total. Through Gene Ontology (GO) and Protein-Protein interaction (PPI) network analysis, we revealed the function and network of these proteins. In summary, we generated replication-competent recombinant influenza viruses by inserting a Strep-Tag into PB2 and purified host factors interacting with viral RdRp bearing a 627 K or 627E PB2. These proteins might function as host range and virulence determinants of influenza virus.

Research Area(s)

  • Host factors, Influenza virus, PB2 627, Protein interaction, Strep-tag viruses

Citation Format(s)

Generation of recombinant influenza virus bearing strep tagged PB2 and effective identification of interactional host factors. / Li, Huanan; Huang, Lihong; Yu, Yuandi; Ren, Xingxing; Li, Bo; Zhang, Jiahao; Liao, Ming; Qi, Wenbao.

In: Veterinary Microbiology, Vol. 254, 108985, 03.2021.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review