Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus

Newcastle disease virus (NDV) genotype VI from pigeon origin is an important causative agent for serious disease in pigeons. Although the biological characteristics of genotype VI NDV have been extensively studied, the understanding of the thermostability of this genotype is still incomplete. In thi...

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Main Authors: Tao Di, Ran Zhao, Qiankai Shi, Fangfang Wang, Zongxi Han, Huixin Li, Yuhao Shao, Junfeng Sun, Shengwang Liu
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/17/7/977
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author Tao Di
Ran Zhao
Qiankai Shi
Fangfang Wang
Zongxi Han
Huixin Li
Yuhao Shao
Junfeng Sun
Shengwang Liu
author_facet Tao Di
Ran Zhao
Qiankai Shi
Fangfang Wang
Zongxi Han
Huixin Li
Yuhao Shao
Junfeng Sun
Shengwang Liu
author_sort Tao Di
collection DOAJ
description Newcastle disease virus (NDV) genotype VI from pigeon origin is an important causative agent for serious disease in pigeons. Although the biological characteristics of genotype VI NDV have been extensively studied, the understanding of the thermostability of this genotype is still incomplete. In this study, an NDV strain, designated P0506, was isolated from a diseased pigeon in China and classified as genotype VI. Phylogenetic analysis on the basis of the Fusion gene coding sequence indicated that P0506 belonged to sub-genotype VI.2.1.1.2.2 of class II. The thermostability may be a universal characteristic of genotype VI NDV. Thus, the thermostability of two strains, including P0506 identified in this study and P0713 identified previously, belonging to VI.2.1.1.2.2, and another previously isolated strain, P0813, in VI.2.1.1.2.1, was investigated. It was indicated that all three viruses presented resistance to heat treatment, but P0713 was more robust than P0813 and P0506. By constructing a series of HN protein mutants, amino acid residues at both residues 365 and 497 in HN protein were found to be involved in the heat resistance. Furthermore, the effects of residues 365 and 497 in HN protein on the thermostability of the virus were further evaluated by using recombinant viruses generated by the reverse genetic system. Our results showed that residue at position 365 in HN protein was the key thermostable determinant of sub-genotype VI.2.1.1.2.2 NDV. These findings will help us better understand the thermostable mechanism of NDV and serve as a foundation for the further development of novel thermostable vaccines.
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spelling doaj-art-bf494c3562664e1abfb201096e0ffdea2025-08-20T03:14:00ZengMDPI AGViruses1999-49152025-07-0117797710.3390/v17070977Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease VirusTao Di0Ran Zhao1Qiankai Shi2Fangfang Wang3Zongxi Han4Huixin Li5Yuhao Shao6Junfeng Sun7Shengwang Liu8State Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaNewcastle disease virus (NDV) genotype VI from pigeon origin is an important causative agent for serious disease in pigeons. Although the biological characteristics of genotype VI NDV have been extensively studied, the understanding of the thermostability of this genotype is still incomplete. In this study, an NDV strain, designated P0506, was isolated from a diseased pigeon in China and classified as genotype VI. Phylogenetic analysis on the basis of the Fusion gene coding sequence indicated that P0506 belonged to sub-genotype VI.2.1.1.2.2 of class II. The thermostability may be a universal characteristic of genotype VI NDV. Thus, the thermostability of two strains, including P0506 identified in this study and P0713 identified previously, belonging to VI.2.1.1.2.2, and another previously isolated strain, P0813, in VI.2.1.1.2.1, was investigated. It was indicated that all three viruses presented resistance to heat treatment, but P0713 was more robust than P0813 and P0506. By constructing a series of HN protein mutants, amino acid residues at both residues 365 and 497 in HN protein were found to be involved in the heat resistance. Furthermore, the effects of residues 365 and 497 in HN protein on the thermostability of the virus were further evaluated by using recombinant viruses generated by the reverse genetic system. Our results showed that residue at position 365 in HN protein was the key thermostable determinant of sub-genotype VI.2.1.1.2.2 NDV. These findings will help us better understand the thermostable mechanism of NDV and serve as a foundation for the further development of novel thermostable vaccines.https://www.mdpi.com/1999-4915/17/7/977Newcastle disease virusgenotype VIhemagglutinin–neuraminidasethermostability
spellingShingle Tao Di
Ran Zhao
Qiankai Shi
Fangfang Wang
Zongxi Han
Huixin Li
Yuhao Shao
Junfeng Sun
Shengwang Liu
Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus
Viruses
Newcastle disease virus
genotype VI
hemagglutinin–neuraminidase
thermostability
title Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus
title_full Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus
title_fullStr Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus
title_full_unstemmed Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus
title_short Residue 365 in Hemagglutinin–Neuraminidase Is a Key Thermostable Determinant of Genotype VI.2.1.1.2.2 Newcastle Disease Virus
title_sort residue 365 in hemagglutinin neuraminidase is a key thermostable determinant of genotype vi 2 1 1 2 2 newcastle disease virus
topic Newcastle disease virus
genotype VI
hemagglutinin–neuraminidase
thermostability
url https://www.mdpi.com/1999-4915/17/7/977
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