Two nuclear localization signals regulate intracellular localization of the duck enteritis virus UL13 protein

Abstract Background UL13 multifunctional tegument protein duck enteritis virus (DEV) is predicted as conserved herpesvirus protein kinase (CHPK); however, little is known about its subcellular localization signal. Results In this study, by transfection with two predicted nuclear signals of DEV UL13 fused to enhanced green fluorescent protein (EGFP), two bipartite nuclear localization signals (NLS) were identified. We found that the NLSs block its nuclear import using ivermectin and proved that nuclear localization signal of DEV UL13 is a classical importin α/β-dependent process. And we constructed the DEV UL13 mutant strain, with the NLSs of DEV UL13 deleted, to explore whether it can affect the virus replication Conclusions The DEV pUL13 amino acids 4 to 7 and 90 to 96 was predicted, and proved that this nuclear import occurs via the classical importin α/β-dependent process. We also found NLSs of pUL13 have no effect on DEV replication in cell culture. Our study enhances the understanding of DEV pUL13. Taken together, these results would provide significant information for the biological function of pUL13 during DEV infection.

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Two nuclear localization signals regulate intracellular localization of the Duck enteritis virus UL13 protein

10.21203/rs.3.rs-15523/v1 ◽

2020 ◽

Author(s):

Linjiang Yang ◽

Xixia Hu ◽

Anchun Cheng ◽

Mingshu Wang ◽

Renyong Jia ◽

...

Keyword(s):

Nuclear Localization ◽

Fluorescent Protein ◽

Intracellular Localization ◽

Duck Enteritis Virus ◽

Significant Information ◽

Nuclear Localization Signals ◽

Dependent Process ◽

Importin Α ◽

Localization Signal ◽

Enteritis Virus

Abstract Background UL13 multifunctional tegument protein of duck enteritis virus (DEV) is predicted as protein kinase (CHPK); however, little is known concerning its subcellular localization signal. Results In this study, by transfection with two predicted nuclear signals of DEV UL13 fused to enhanced green fluorescent protein (EGFP), two bipartite nuclear localization signal (NLS) was identified. We identified the nuclear localization signals (NLSs) that control its nuclear importing using fluorescence assay and proved that nuclear localization of DEV UL13 is a classical importin α/β-dependent process. And we constructed the mutant DEV, with the NLSs of UL13 deleted, to explore whether it will affect the replication of virus particles. Conclusions DEV UL13 protein is directed by amino acids 4 to 7 and 90 to 96, and proved that this nuclear import occurs via the classical importin α/β-dependent process. And Entry nucleus of UL13 protein has no effect on DEV replication in cell culture. Our study enhances the understanding of DEV pUL13. Taken together, these results would provide significant information for the stud y of the biological function of UL13 during DEV infection.

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Identification of the Nuclear Localization Signal Region of Duck Enteritis Virus UL14 and Its Interaction with VP16

Intervirology ◽

10.1159/000452711 ◽

2016 ◽

Vol 59 (4) ◽

pp. 187-196

Author(s):

FangJie Li ◽

YuWei Zhang ◽

Shun Chen ◽

MingShu Wang ◽

RenYong Jia ◽

...

Keyword(s):

Nuclear Localization ◽

Nuclear Localization Signal ◽

Duck Enteritis Virus ◽

Signal Region ◽

Localization Signal ◽

Enteritis Virus

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Characterization of nucleocytoplasmic shuttling and intracellular localization signals in Duck Enteritis Virus UL54

Biochimie ◽

10.1016/j.biochi.2016.05.003 ◽

2016 ◽

Vol 127 ◽

pp. 86-94 ◽

Cited By ~ 8

Author(s):

Chaoyue Liu ◽

Anchun Cheng ◽

Mingshu Wang ◽

Shun Chen ◽

Renyong Jia ◽

...

Keyword(s):

Intracellular Localization ◽

Duck Enteritis Virus ◽

Nucleocytoplasmic Shuttling ◽

Enteritis Virus

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Duck enteritis virus pUL47, as a late structural protein localized in the nucleus, mainly depends on residues 40 to 50 and 768 to 777 and inhibits IFN-β signalling by interacting with STAT1

Veterinary Research ◽

10.1186/s13567-020-00859-w ◽

2020 ◽

Vol 51 (1) ◽

Author(s):

Tianqiong He ◽

Mingshu Wang ◽

Anchun Cheng ◽

Qiao Yang ◽

Renyong Jia ◽

...

Keyword(s):

Nuclear Localization ◽

Fluorescent Protein ◽

Nuclear Translocation ◽

Duck Enteritis Virus ◽

Poly I:C ◽

Nuclear Localization Sequence ◽

Heterologous Proteins ◽

Structural Protein ◽

Oligoadenylate Synthetase ◽

Enteritis Virus

Abstract Duck enteritis virus (DEV) is a member of the Alphaherpesvirinae subfamily. The characteristics of some DEV genes have been reported. However, information regarding the DEV UL47 gene is limited. In this study, we identified the DEV UL47 gene encoding a late structural protein located in the nucleus of infected cells. We further found that two domains of DEV pUL47, amino acids (aa) 40 to 50 and 768 to 777, could function as nuclear localization sequence (NLS) to guide the nuclear localization of pUL47 and nuclear translocation of heterologous proteins, including enhanced green fluorescent protein (EGFP) and beta-galactosidase (β-Gal). Moreover, pUL47 significantly inhibited polyriboinosinic:polyribocytidylic acid [poly(I:C)]-induced interferon beta (IFN-β) production and downregulated interferon-stimulated gene (ISG) expression, such as Mx and oligoadenylate synthetase-like (OASL), by interacting with signal transducer and activator of transcription-1 (STAT1).

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Expression and intracellular localization of duck enteritis virus pUL38 protein

Virology Journal ◽

10.1186/1743-422x-7-162 ◽

2010 ◽

Vol 7 (1) ◽

pp. 162 ◽

Cited By ~ 12

Author(s):

Jun Xiang ◽

Guangpeng Ma ◽

Shunchuan Zhang ◽

Anchun Cheng ◽

Mingshu Wang ◽

...

Keyword(s):

Intracellular Localization ◽

Duck Enteritis Virus ◽

Enteritis Virus

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Proteomic analysis of host cellular proteins co-immunoprecipitated with duck enteritis virus gC

Journal of Proteomics ◽

10.1016/j.jprot.2021.104281 ◽

2021 ◽

Vol 245 ◽

pp. 104281

Author(s):

Liu Chen ◽

Zheng Ni ◽

Jionggang Hua ◽

Weicheng Ye ◽

Keshu Liu ◽

...

Keyword(s):

Proteomic Analysis ◽

Duck Enteritis Virus ◽

Cellular Proteins ◽

Enteritis Virus

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Development of a Colloidal Gold Immunochromatographic Assay for Duck Enteritis Virus Detection Using Monoclonal Antibodies

Pathogens ◽

10.3390/pathogens10030365 ◽

2021 ◽

Vol 10 (3) ◽

pp. 365

Author(s):

Fengli Liu ◽

Yanxin Cao ◽

Maokai Yan ◽

Mengxu Sun ◽

Qingshui Zhang ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Colloidal Gold ◽

Preventive Intervention ◽

Virus Detection ◽

Cross Reactivity ◽

Duck Enteritis Virus ◽

Detection Sensitivity ◽

Immunochromatographic Assay ◽

Efficient Detection ◽

Enteritis Virus

Duck viral enteritis is a highly contagious and fatal disease of commercial waterfowl flocks. The disease occurs sporadically or epizootically in mainland China due to insufficient vaccinations. Early and rapid diagnosis is important for preventive intervention and the control of epizootic events in clinical settings. In this study, we generated two monoclonal antibodies (MAbs) that specifically recognized the duck enteritis virus (DEV) envelope glycoprotein B and tegument protein UL47, respectively. Using these MAbs, a colloidal gold-based immunochromatographic assay (ICA) was developed for the efficient detection of DEV antigens within 15 min. Our results showed that the detection limit of the developed ICA strip was 2.52 × 103 TCID50/mL for the virus infected cell culture suspension with no cross-reactivity with other pathogenic viruses commonly encountered in commercially raised waterfowl. Using samples from experimentally infected ducks, we demonstrated that the ICA detected the virus in cloacal swab samples on day three post-infection, demonstrating an 80% concordance with the PCR. For tissue homogenates from ducks succumbing to infection, the detection sensitivity was 100%. The efficient and specific detection by this ICA test provides a valuable, convenient, easy to use and rapid diagnostic tool for DVE under both laboratory and field conditions.

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