Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts

Autor: Houqiang Xu, Xinqin Ji, Jiafu Zhao, Xiufan Liu, Haixu Xu, Hu Yan, Shunlin Hu, Deng Shanshan, Zhiqiang Duan
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Microbiology (medical)
animal structures
Newcastle Disease
viruses
Immunology
Active Transport
Cell Nucleus
Newcastle disease virus
matrix protein
Importin
Biology
Karyopherins
Recombinant virus
Virus Replication
Microbiology
Models
Biological
environment and public health
Cell Line
lcsh:Infectious and parasitic diseases
Viral Matrix Proteins
03 medical and health sciences
Two-Hybrid System Techniques
Protein Interaction Mapping
NLS
Animals
Immunoprecipitation
lcsh:RC109-216
chicken fibroblasts
nuclear import mechanism
Viral matrix protein
Fibroblasts
nuclear localization signal
Cell biology
030104 developmental biology
Infectious Diseases
Viral replication
Gene Expression Regulation
Ran
embryonic structures
Parasitology
Nuclear transport
Chickens
Nuclear localization sequence
Research Paper
Zdroj: Virulence, Vol 9, Iss 1, Pp 783-803 (2018)
Virulence
ISSN: 2150-5608
2150-5594
Popis: The matrix (M) protein of Newcastle disease virus (NDV) is demonstrated to localize in the nucleus via intrinsic nuclear localization signal (NLS), but cellular proteins involved in the nuclear import of NDV M protein and the role of M's nuclear localization in the replication and pathogenicity of NDV remain unclear. In this study, importin β1 was screened to interact with NDV M protein by yeast two-hybrid screening. This interaction was subsequently confirmed by co-immunoprecipitation and pull-down assays. In vitro binding studies indicated that the NLS region of M protein and the amino acids 336–433 of importin β1 that belonged to the RanGTP binding region were important for binding. Importantly, a recombinant virus with M/NLS mutation resulted in a pathotype change of NDV and attenuated viral replication and pathogenicity in chicken fibroblasts and SPF chickens. In agreement with the binding data, nuclear import of NDV M protein in digitonin-permeabilized HeLa cells required both importin β1 and RanGTP. Interestingly, importin α5 was verified to interact with M protein through binding importin β1. However, importin β1 or importin α5 depletion by siRNA resulted in different results, which showed the obviously cytoplasmic or nuclear accumulation of M protein and the remarkably decreased or increased replication ability and pathogenicity of NDV in chicken fibroblasts, respectively. Our findings therefore demonstrate for the first time the nuclear import mechanism of NDV M protein and the negative regulation role of importin α5 in importin β1-mediated nuclear import of M protein and the replication and pathogenicity of a paramyxovirus.
Databáze: OpenAIRE
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