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Biosci Trends. 2019 Jul 22;13(3):225-233. doi: 10.5582/bst.2019.01070. Epub 2019 May 28.

Conserved amino acids around the DIII-DI linker region of the Newcastle disease virus fusion protein are critical for protein folding and fusion activity.

Author information

1
Department of Virology, School of Public Health, Shandong University.
2
Department of Laboratory Medicine, Shandong Provincial Qianfoshan Hospital, Shandong University.
3
State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University.
4
The Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University.

Abstract

Newcastle disease virus (NDV), an avian paramyxovirus, causes Newcastle disease (ND) which is a highly contagious and fatal viral disease affecting poultry and most species of birds. The fusion (F) protein of NDV mediates membrane fusion, which is essential to the processes of viral entry, replication, and dissemination. Although several domains of NDV F are known to have important effects on regulating the membrane fusion activity, the role of the region around domain III (DIII) and domain I (DI) still remains ill-defined. Site-directed mutagenesis was utilized to change the conserved amino acids at 269, 274, 277, 286, 287, 290, 295, and 297 to alanine in order to investigate the effects of these conserved amino acids around the DIII and DI linker region of the NDV F protein on fusion activity. It was found that five of these substitutions almost abolished fusion activity except for mutants I269A, Q286A, and N297A, which showed 57.1%, 161.1%, and 97.7% of the wt F level, respectively. Four (I274A, D277A, V287A, and P290A) of these five mutants likely result in interfering with folding or transporting of the molecule since these proteins were minimally expressed at the cell surface, formed aggregates, or not proteolytically cleaved. However, mutant L295A almost abolished fusion activity even with a similar level of cell surface expression. These data indicated that conserved amino acids around the DIII-DI linker region are critical for the folding of the F protein and have an important influence on fusion activity.

KEYWORDS:

F protein; Membrane fusion; NDV; Protein folding and transport

PMID:
31142702
DOI:
10.5582/bst.2019.01070
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