Duck plague virus gE serves essential functions during the virion final envelopment through influence capsids budding into the cytoplasmic vesicles

Tian Liu,Mujeeb Ur Rehman,Xiaoyue Chen,Xinxin Zhao,Leichang Pan,Shun Chen,Mingshu Wang,Dekang Zhu,Ling Zhang,Shaqiu Zhang,Yunya Liu,Qiao Yang,Anchun Cheng,Renyong Jia,Mafeng Liu,Ying Wu,Yanling Yu,Bin Tian

doi:10.1038/s41598-020-62604-9

Tian Liu, Mujeeb Ur Rehman + Show 16 more

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Journal: Scientific Reports	Publication Date: Mar 27, 2020
Citations: 13	License type: open-access

Affiliation: Sichuan Agricultural University

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Duck plague virus (DPV), a member of the alphaherpesviruses subfamily, causes massive ducks death and results in a devastating hit to duck industries in China. It is of great significance for us to analyze the functions of DPV genes for controlling the outbreak of duck plague. Thus, glycoproteins E (gE) of DPV, which requires viral cell-to-cell spreading and the final envelopment in herpes simplex virus 1 (HSV-1) and pseudorabies virus (PRV), was chosen herein. The gE mutant virus BAC-CHv-ΔgE was constructed by using a markerless two-step Red recombination system implemented on the DPV genome cloned into a bacterial artificial chromosome (BAC). Viral plaques on duck embryo fibroblast (DEF) cells of BAC-CHv-ΔgE were on average approximately 60% smaller than those produced by BAC-CHv virus. Viral replication kinetics showed that BAC-CHv-ΔgE grew to lower titers than BAC-CHv virus did in DEF cells. Electron microscopy confirmed that deleting of DPV gE resulted in a large number of capsids accumulating around vesicles and very few of them could bud into vesicles. The drastic inhibition of virion formation in the absence of the DPV gE indicated that it played an important role in virion morphogenesis before the final envelopment of intracytoplasmic nucleocapsids.

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Duck plague virus (DPV), a member of the alphaherpesviruses subfamily, causes massive ducks death and results in a devastating hit to duck industries in China
To explore the functions of DPV glycoproteins E (gE) in-depth, gE gene was deleted by using a markerless two-step Red recombination system implemented on the DPV genome that was cloned into a bacterial artificial chromosome (BAC)[17]
To characterize the functions of DPV gE, we successfully constructed a mutant derived from a BAC copy containing the Chinese virulent strain (CHv) strain genome in which the gE coding sequence was deleted by a markerless two-step Red recombination system; the mutant was named BAC-CHv-ΔgE (Fig. 1a–c)

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Duck plague virus (DPV), a member of the alphaherpesviruses subfamily, causes massive ducks death and results in a devastating hit to duck industries in China It is of great significance for us to analyze the functions of DPV genes for controlling the outbreak of duck plague. In PRV, extracellular and intracytoplasmic enveloped virus particles are absent in the absence of gE and gM genes, and unenveloped capsids accumulate in the cytoplasm surrounded by tegument proteins in infected cells[16]. Electron microscopy analysis confirmed that deleting DPV gE impaired the final viral envelopment, during which fewer capsids could bud into cytoplasmic vesicles.

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