1. Mutations in the NDV fusion protein HR4 region decreased fusogenic activity due to failed protein expression.
- Author
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Liu Y, Pan H, Wang H, Yuan Y, Cao F, Song W, and Wang Z
- Subjects
- Animals, Amino Acid Substitution, Cell Line, Mutation, Proteolysis, Membrane Fusion, Newcastle disease virus genetics, Newcastle disease virus metabolism, Viral Fusion Proteins genetics, Viral Fusion Proteins metabolism, Virus Internalization, Mutagenesis, Site-Directed
- Abstract
Newcastle disease virus (NDV) is the pathogen of a zoonosis that is primarily transmitted by poultry and has severe infectivity and a high fatality rate. Many studies have focused on the role of the NDV fusion (F) protein in the cell-cell membrane fusion process. However, little attention has been given to the heptad repeat region, HR4, which is located in the NDV F2 subunit. Here, site-directed mutants were constructed to study the function of the NDV F protein HR4 region and identify the key amino acids in this region. Nine conserved amino acids were substituted with alanine or the corresponding amino acid of other aligned paramyxoviruses. The desired mutants were examined for changes in fusogenic activity through three kinds of membrane fusion assays and expression and proteolysis through IFA, FACS and WB. The results showed that when conserved amino acids (L81, Y84, L88, L91, L92, P94, L95 and I99) were replaced with alanine, the fusogenic activity of the F protein was abolished, possibly because of failed protein expression not only on the cell surface but also inside cells. These data indicated that the conserved amino acids above in NDV F HR4 are critical for normal protein synthesis and expression, possibly for the stability of the F protein monomer, formation of trimer and conformational changes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Zhiyu Wang reports financial support was provided by National Natural Science Foundation of China. Yaqing Liu reports financial support was provided by Doctoral Research Foundation of Affiliated Hospital of Qingdao University. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
- Published
- 2024
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