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Porcine epidemic diarrhea virus (PEDV) is a highly pathogenic porcine enteropathogenic coronavirus causing severe enteritis and lethal watery diarrhea in piglets. PEDV infection suppresses the synthesis of type I IFN, and multiple viral proteins of PEDV have been shown to target the adaptors of innate immune pathways to inhibit type I IFN production. In this study, we identified PEDV membrane (M) protein as a new antagonist of type I IFN production in both human embryonic kidney HEK293T cells and porcine kidney PK-15 cells and determined the antagonistic mechanism used by M protein to target IFN regulatory factor 7 (IRF7), an important regulator of type I IFN production. IRF7 is phosphorylated and activated by TBK1 and IKKε in response to viral infection. We found that PEDV M protein interacted with the inhibitory domain of IRF7 and significantly suppressed TBK1/IKKε-induced IRF7 phosphorylation and dimerization of IRF7, leading to the decreased expression of type I IFN, although it did not affect the interaction between TBK1/IKKε and IRF7. As expected, overexpression of M protein significantly increased PEDV replication in porcine cells. The M proteins of both epidemic PEDV strains and vaccine strain showed similar antagonistic effect on type I IFN production, and the 1-55 region of M protein was essential for disruption of IRF7 function by interacting with IRF7. Taken together, our data identified a new, to our knowledge, IFN antagonist of PEDV, as well as a novel, to our knowledge, antagonistic mechanism evolved by PEDV to inhibit type I IFN production. Key Points PEDV M protein inhibits type I IFN production. PEDV M protein interacts with the ID of IRF7 to suppress IRF7 activation. The 1-55 region of M protein is essential for disruption of IRF7 function.
Shasha Li; Zixiang Zhu; Fan Yang; Weijun Cao; Jinping Yang; Caina Ma; Zhenxiang Zhao; Hong Tian; Xiangtao Liu; Junwu Ma; Shaobo Xiao; Haixue Zheng. Porcine Epidemic Diarrhea Virus Membrane Protein Interacted with IRF7 to Inhibit Type I IFN Production during Viral Infection. The Journal of Immunology 2021, 206, ji2001186 -2923.
AMA StyleShasha Li, Zixiang Zhu, Fan Yang, Weijun Cao, Jinping Yang, Caina Ma, Zhenxiang Zhao, Hong Tian, Xiangtao Liu, Junwu Ma, Shaobo Xiao, Haixue Zheng. Porcine Epidemic Diarrhea Virus Membrane Protein Interacted with IRF7 to Inhibit Type I IFN Production during Viral Infection. The Journal of Immunology. 2021; 206 (12):ji2001186-2923.
Chicago/Turabian StyleShasha Li; Zixiang Zhu; Fan Yang; Weijun Cao; Jinping Yang; Caina Ma; Zhenxiang Zhao; Hong Tian; Xiangtao Liu; Junwu Ma; Shaobo Xiao; Haixue Zheng. 2021. "Porcine Epidemic Diarrhea Virus Membrane Protein Interacted with IRF7 to Inhibit Type I IFN Production during Viral Infection." The Journal of Immunology 206, no. 12: ji2001186-2923.
Coronavirus accessory proteins are a unique set of proteins whose genes are interspersed among or within the genes encoding structural proteins. Different coronavirus genera, or even different species within the same coronavirus genus, encode varying amounts of accessory proteins, leading to genus- or species-specificity. Though accessory proteins are dispensable for the replication of coronavirus in vitro, they play important roles in regulating innate immunity, viral proliferation, and pathogenicity. The function of accessory proteins on virus infection and pathogenesis is an area of particular interest. In this review, we summarize the current knowledge on accessory proteins of several representative coronaviruses that infect humans or animals, including the emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with an emphasis on their roles in interaction between virus and host, mainly involving stress response, innate immunity, autophagy, and apoptosis. The cross-talking among these pathways is also discussed.
Puxian Fang; Liurong Fang; Huichang Zhang; Sijin Xia; Shaobo Xiao. Functions of Coronavirus Accessory Proteins: Overview of the State of the Art. Viruses 2021, 13, 1139 .
AMA StylePuxian Fang, Liurong Fang, Huichang Zhang, Sijin Xia, Shaobo Xiao. Functions of Coronavirus Accessory Proteins: Overview of the State of the Art. Viruses. 2021; 13 (6):1139.
Chicago/Turabian StylePuxian Fang; Liurong Fang; Huichang Zhang; Sijin Xia; Shaobo Xiao. 2021. "Functions of Coronavirus Accessory Proteins: Overview of the State of the Art." Viruses 13, no. 6: 1139.
This is the first study to clone duck CCCH-type zinc finger antiviral protein (duZAP) from Jingjiang duck (Anas platyrhynchos). Full-length duZAP cDNA was 2154 bp and encoded a 717-amino acid polypeptide containing four highly conserved CCCH-type finger motifs, a WWE domain and a poly (ADP-ribose) polymerase (PARP) domain. duZAP was expressed in multiple duck tissues, with the highest mRNA expression in the spleen. Overexpression of duZAP in duck embryo fibroblast cells (DEFs) led to activation of the transcription factors IRF1 and NF-κB, and induction of IFN-β. Analysis of deletion mutants revealed that both the WWE and PARP domains of duZAP were essential for activating the IFN-β promoter. Knockdown of duZAP in DEFs significantly reduced poly (I:C)- and duck Tembusu virus (DTMUV)-induced IFN-β activation. Our findings further the understanding of the role of duZAP in the duck innate immune response.
Rongrong Zhang; Yan He; Xinyu Zhu; Guoyuan Wen; Qingping Luo; Tengfei Zhang; Qin Lu; Shudan Liu; Shaobo Xiao; Liurong Fang; Huabin Shao. Molecular characterization and functional analysis of duck CCCH-type zinc finger antiviral protein (ZAP). Biochemical and Biophysical Research Communications 2021, 561, 52 -58.
AMA StyleRongrong Zhang, Yan He, Xinyu Zhu, Guoyuan Wen, Qingping Luo, Tengfei Zhang, Qin Lu, Shudan Liu, Shaobo Xiao, Liurong Fang, Huabin Shao. Molecular characterization and functional analysis of duck CCCH-type zinc finger antiviral protein (ZAP). Biochemical and Biophysical Research Communications. 2021; 561 ():52-58.
Chicago/Turabian StyleRongrong Zhang; Yan He; Xinyu Zhu; Guoyuan Wen; Qingping Luo; Tengfei Zhang; Qin Lu; Shudan Liu; Shaobo Xiao; Liurong Fang; Huabin Shao. 2021. "Molecular characterization and functional analysis of duck CCCH-type zinc finger antiviral protein (ZAP)." Biochemical and Biophysical Research Communications 561, no. : 52-58.
Porcine epidemic diarrhea virus (PEDV) is a reemerging Alphacoronavirus that causes lethal diarrhea in piglets. Coronavirus nonstructural protein 13 (nsp13) encodes helicase, which plays pivotal roles during viral replication by unwinding viral RNA. However, the biochemical characterization of PEDV nsp13 remains largely unknown. In this study, PEDV nsp13 was expressed in Escherichia coli and purified. The recombinant nsp13 possessed ATPase and helicase activities for binding and unwinding dsDNA/RNA substrates with 5′-overhangs, and Mg2+ and Mn2+ were critical for its ATPase and helicase activities. PEDV nsp13 also unwound dsDNA into ssDNA in the pH from 6.0–9.0, and used energy from all nucleoside triphosphates and deoxynucleoside triphosphates. Site-directed mutagenesis demonstrated that Lys289 (K289) of PEDV nsp13 was essential for its ATPase and helicase activities. These results provide new insights into the biochemical properties of PEDV nsp13, which is a potential target for developing antiviral drugs.
Jie Ren; Zhen Ding; Puxian Fang; Shaobo Xiao; Liurong Fang. ATPase and helicase activities of porcine epidemic diarrhea virus nsp13. Veterinary Microbiology 2021, 257, 109074 .
AMA StyleJie Ren, Zhen Ding, Puxian Fang, Shaobo Xiao, Liurong Fang. ATPase and helicase activities of porcine epidemic diarrhea virus nsp13. Veterinary Microbiology. 2021; 257 ():109074.
Chicago/Turabian StyleJie Ren; Zhen Ding; Puxian Fang; Shaobo Xiao; Liurong Fang. 2021. "ATPase and helicase activities of porcine epidemic diarrhea virus nsp13." Veterinary Microbiology 257, no. : 109074.
Porcine deltacoronavirus (PDCoV) is a novel swine enteropathogenic coronavirus that causes serious vomiting and diarrhea in piglets. Previous work demonstrated that PDCoV infection inhibits type I interferon (IFN) production. Here, we found that ectopic expression of PDCoV nsp10 significantly inhibited Sendai virus (SeV)-induced IFN-β production by impairing the phosphorylation and nuclear translocation of two transcription factors, IRF3 and NF-κB p65 subunit. Interestingly, experiments with truncated mutants and site-directed mutagenesis revealed that PDCoV nsp10 mutants with missing or destroyed zinc fingers (ZFs) domains also impeded SeV-induced IFN-β production, suggesting that nsp10 does not require its ZF domains to antagonize IFN-β production. Further work found that co-expression of nsp10 with nsp14 or nsp16, two replicative enzymes, significantly enhanced the inhibitory effects of nsp10 on IFN-β. Taken together, our results demonstrate that PDCoV nsp10 antagonizes IFN via a ZF-independent mechanism and has a synergistic effect with nsp14 and nsp16 on inhibiting IFN-β production.
Puxian Fang; Yingying Hong; Sijin Xia; Jiansong Zhang; Jie Ren; Yanrong Zhou; Liurong Fang; Shaobo Xiao. Porcine deltacoronavirus nsp10 antagonizes interferon-β production independently of its zinc finger domains. Virology 2021, 559, 46 -56.
AMA StylePuxian Fang, Yingying Hong, Sijin Xia, Jiansong Zhang, Jie Ren, Yanrong Zhou, Liurong Fang, Shaobo Xiao. Porcine deltacoronavirus nsp10 antagonizes interferon-β production independently of its zinc finger domains. Virology. 2021; 559 ():46-56.
Chicago/Turabian StylePuxian Fang; Yingying Hong; Sijin Xia; Jiansong Zhang; Jie Ren; Yanrong Zhou; Liurong Fang; Shaobo Xiao. 2021. "Porcine deltacoronavirus nsp10 antagonizes interferon-β production independently of its zinc finger domains." Virology 559, no. : 46-56.
Outbreaks of coronaviruses, especially SARS-CoV-2, pose a serious threat to global public health. Development of vaccines to prevent the coronaviruses that can infect humans has always been a top priority.
Yuejun Shi; Jiale Shi; Limeng Sun; Yubei Tan; Gang Wang; Fenglin Guo; Guangli Hu; Yanan Fu; Zhen F. Fu; Shaobo Xiao; Guiqing Peng. Insight into Vaccine Development for Alphacoronaviruses Based on Structural and Immunological Analyses of Spike Proteins. Journal of Virology 2021, 95, 1 .
AMA StyleYuejun Shi, Jiale Shi, Limeng Sun, Yubei Tan, Gang Wang, Fenglin Guo, Guangli Hu, Yanan Fu, Zhen F. Fu, Shaobo Xiao, Guiqing Peng. Insight into Vaccine Development for Alphacoronaviruses Based on Structural and Immunological Analyses of Spike Proteins. Journal of Virology. 2021; 95 (7):1.
Chicago/Turabian StyleYuejun Shi; Jiale Shi; Limeng Sun; Yubei Tan; Gang Wang; Fenglin Guo; Guangli Hu; Yanan Fu; Zhen F. Fu; Shaobo Xiao; Guiqing Peng. 2021. "Insight into Vaccine Development for Alphacoronaviruses Based on Structural and Immunological Analyses of Spike Proteins." Journal of Virology 95, no. 7: 1.
Type I IFNs (IFN-Is) play pivotal roles in host defense against viral infections but remain enigmatic against bacterial pathogens. In this study, we recombinantly expressed and purified intact grass carp (Ctenopharyngodon idella) IFNφ1 (gcIFNφ1), a teleost IFN-I. gcIFNφ1 widely powerfully directly kills both Gram-negative and Gram-positive bacteria in a dose-dependent manner. gcIFNφ1 binds to LPS or peptidoglycan and provokes bacterial membrane depolarization and disruption, resulting in bacterial death. Furthermore, gcIFNφ1 can efficiently protect zebrafish against Aeromonas hydrophila infection and significantly reduce the bacterial loads in tissues by an infection model. In addition, we wonder whether antibacterial IFN-I members exist in other vertebrates. The amino acid compositions of representative IFN-Is with strong positive charges from Pisces, Amphibia, reptiles, Aves, and Mammalia demonstrate high similarities with those of 2237 reported cationic antimicrobial peptides in antimicrobial peptide database. Recombinant intact representative IFN-I members from the nonmammalian sect exhibit potent broad-spectrum robust bactericidal activity through bacterial membrane depolarization; in contrast, the bactericidal activity is very weak from mammalian IFN-Is. The findings display a broad-spectrum potent direct antimicrobial function for IFN-Is, to our knowledge previously unknown. The results highlight that IFN-Is are important and robust in host defense against bacterial pathogens, and unify direct antibacterial and indirect antiviral bifunction in nonmammalian jawed vertebrates. Key Points Teleost IFNφ1 was discovered as a broad powerful bactericidal cytokine. IFNφ1 directly kills bacteria by cytoderm binding and disruption. Potent antimicrobial peptide–like IFN-Is were identified in other gnathostomes.
Xun Xiao; Wentao Zhu; Yanqi Zhang; Zhiwei Liao; Changsong Wu; Chunrong Yang; Yongan Zhang; Shaobo Xiao; Jianguo Su. Broad-Spectrum Robust Direct Bactericidal Activity of Fish IFNφ1 Reveals an Antimicrobial Peptide–like Function for Type I IFNs in Vertebrates. The Journal of Immunology 2021, 206, 1337 -1347.
AMA StyleXun Xiao, Wentao Zhu, Yanqi Zhang, Zhiwei Liao, Changsong Wu, Chunrong Yang, Yongan Zhang, Shaobo Xiao, Jianguo Su. Broad-Spectrum Robust Direct Bactericidal Activity of Fish IFNφ1 Reveals an Antimicrobial Peptide–like Function for Type I IFNs in Vertebrates. The Journal of Immunology. 2021; 206 (6):1337-1347.
Chicago/Turabian StyleXun Xiao; Wentao Zhu; Yanqi Zhang; Zhiwei Liao; Changsong Wu; Chunrong Yang; Yongan Zhang; Shaobo Xiao; Jianguo Su. 2021. "Broad-Spectrum Robust Direct Bactericidal Activity of Fish IFNφ1 Reveals an Antimicrobial Peptide–like Function for Type I IFNs in Vertebrates." The Journal of Immunology 206, no. 6: 1337-1347.
Cholesterol 25-hydroxylase (CH25 H) is a key enzyme regulating cholesterol metabolism and also acts as a broad antiviral host restriction factor. Porcine deltacoronavirus (PDCoV) is an emerging swine enteropathogenic coronavirus that can cause vomiting, diarrhea, dehydration and even death in newborn piglets. In this study, we found that PDCoV infection significantly upregulated the expression of CH25H in IPI-FX cells, a cell line of porcine ileum epithelium. Overexpression of CH25H inhibited PDCoV replication, whereas CH25H silencing using RNA interference promoted PDCoV infection. Treatment with 25-hydroxycholesterol (25HC), the catalysate of cholesterol via CH25H, inhibited PDCoV proliferation by impairing viral invasion of IPI-FX cells. Furthermore, a mutant CH25H (CH25H-M) lacking hydroxylase activity also inhibited PDCoV infection to a lesser extent. Taken together, our data suggest that CH25H acts as a host restriction factor to inhibit the proliferation of PDCoV but this inhibitory effect is not completely dependent on its enzymatic activity.
Wenting Ke; Xiaoli Wu; Puxian Fang; Yanrong Zhou; Liurong Fang; Shaobo Xiao. Cholesterol 25-hydroxylase suppresses porcine deltacoronavirus infection by inhibiting viral entry. Virus Research 2021, 295, 198306 -198306.
AMA StyleWenting Ke, Xiaoli Wu, Puxian Fang, Yanrong Zhou, Liurong Fang, Shaobo Xiao. Cholesterol 25-hydroxylase suppresses porcine deltacoronavirus infection by inhibiting viral entry. Virus Research. 2021; 295 ():198306-198306.
Chicago/Turabian StyleWenting Ke; Xiaoli Wu; Puxian Fang; Yanrong Zhou; Liurong Fang; Shaobo Xiao. 2021. "Cholesterol 25-hydroxylase suppresses porcine deltacoronavirus infection by inhibiting viral entry." Virus Research 295, no. : 198306-198306.
Coronaviruses spike (S) glycoproteins mediate viral entry into host cells by binding to host receptors. However, how the S1 subunit undergoes conformational changes for receptor recognition has not been elucidated inAlphacoronavirus. Here, we report the cryo-EM structures of the HCoV-229E S trimer in prefusion state with two conformations. The activated conformation may pose the potential exposure of the S1-RBDs by decreasing of the interaction area between the S1-RBDs and the surrounding S1-NTDs and S1-RBDs compared to the closed conformation. Furthermore, structural comparison of our structures with the previously reported HCoV-229E S structure showed that the S trimers trended to open the S2 subunit from the closed conformation to open conformation, which could promote the transition from pre- to postfusion. Our results provide insights into the mechanisms involved in S glycoprotein-mediatedAlphacoronavirusentry and have implications for vaccine and therapeutic antibody design.
Xiyong Song; Yuejun Shi; Wei Ding; Tongxin Niu; Limeng Sun; Yubei Tan; Yong Chen; Jiale Shi; QiQi Xiong; Xiaojun Huang; Shaobo Xiao; Yanping Zhu; Chongyun Cheng; Zhen F. Fu; Zhi-Jie Liu; Guiqing Peng. Cryo-EM analysis of the HCoV-229E spike glycoprotein reveals dynamic prefusion conformational changes. Nature Communications 2021, 12, 1 -9.
AMA StyleXiyong Song, Yuejun Shi, Wei Ding, Tongxin Niu, Limeng Sun, Yubei Tan, Yong Chen, Jiale Shi, QiQi Xiong, Xiaojun Huang, Shaobo Xiao, Yanping Zhu, Chongyun Cheng, Zhen F. Fu, Zhi-Jie Liu, Guiqing Peng. Cryo-EM analysis of the HCoV-229E spike glycoprotein reveals dynamic prefusion conformational changes. Nature Communications. 2021; 12 (1):1-9.
Chicago/Turabian StyleXiyong Song; Yuejun Shi; Wei Ding; Tongxin Niu; Limeng Sun; Yubei Tan; Yong Chen; Jiale Shi; QiQi Xiong; Xiaojun Huang; Shaobo Xiao; Yanping Zhu; Chongyun Cheng; Zhen F. Fu; Zhi-Jie Liu; Guiqing Peng. 2021. "Cryo-EM analysis of the HCoV-229E spike glycoprotein reveals dynamic prefusion conformational changes." Nature Communications 12, no. 1: 1-9.
Coronaviruses (CoVs) are potential pandemic pathogens that can infect a variety of hosts and cause respiratory, enteric, hepatic and neurological diseases. Nonstructural protein 3 (nsp3), an essential component of the replication/transcription complex, is one of the most important antiviral targets. Here, we report the first crystal structure of multiple functional domains from porcine delta-coronavirus (PDCoV) nsp3, including the macro domain (Macro), ubiquitin-like domain 2 (Ubl2) and papain-like protease (PLpro) catalytic domain. In the asymmetric unit, two of the subunits form the head-to-tail homodimer with an interaction interface between Macro and PLpro. However, PDCoV Macro-Ubl2-PLpro mainly exists as a monomer in solution. Then, we conducted fluorescent resonance energy transfer-based protease assays and found that PDCoV PLpro can cleave a peptide by mimicking the cognate nsp2/nsp3 cleavage site in peptide substrates and exhibits deubiquitinating and de-interferon stimulated gene(deISGylating) activities by hydrolysing ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) and ISG15-AMC substrates. Moreover, the deletion of Macro or Macro-Ubl2 decreased the enzyme activity of PLpro, indicating that Macro and Ubl2 play important roles in maintaining the stability of the PLpro domain. Two active sites of PLpro, Cys260 and His398, were determined; unexpectedly, the conserved site Asp412 was not the third active site. Furthermore, the motif “NGYDT” (amino acids 409–413) was important for stabilizing the enzyme activity of PLpro, and the N409A mutant significantly decreased the enzyme activity of PLpro. These results provide novel insights into the replication mechanism of CoV and new clues for future drug design.
Mengxia Li; Gang Ye; Yu Si; Zhou Shen; Zhu Liu; Yuejun Shi; Shaobo Xiao; Zhen F. Fu; Guiqing Peng. Structure of the multiple functional domains from coronavirus nonstructural protein 3. Emerging Microbes & Infections 2021, 10, 66 -80.
AMA StyleMengxia Li, Gang Ye, Yu Si, Zhou Shen, Zhu Liu, Yuejun Shi, Shaobo Xiao, Zhen F. Fu, Guiqing Peng. Structure of the multiple functional domains from coronavirus nonstructural protein 3. Emerging Microbes & Infections. 2021; 10 (1):66-80.
Chicago/Turabian StyleMengxia Li; Gang Ye; Yu Si; Zhou Shen; Zhu Liu; Yuejun Shi; Shaobo Xiao; Zhen F. Fu; Guiqing Peng. 2021. "Structure of the multiple functional domains from coronavirus nonstructural protein 3." Emerging Microbes & Infections 10, no. 1: 66-80.
Currently, the active-site residues and substrate specificities of 3C-like protease (3CL pro ) differ among nidoviruses, and the detailed catalytic mechanism remains largely unknown. Here, porcine torovirus (PToV) 3CL pro cleaves 12 sites in the polyproteins, including its N- and C-terminal self-processing sites. Unlike coronaviruses and arteriviruses, PToV 3CL pro employed His53 and Ser160 as the active-site residues that recognize a glutamine (Gln) at the P1 position. Surprisingly, mutations of P1-Gln impaired the C-terminal self-processing but did not affect N-terminal self-processing. The “noncanonical” substrate specificity for its N-terminal self-processing was attributed to the phenylalanine (Phe) residue at the P4 position in the N-terminal site. Furthermore, a double glycine (neutral) substitution at the putative P4-Phe-binding residues (P62G/L185G) abolished the cleavage activity of PToV 3CL pro suggested the potential hydrophobic force between the PToV 3CL pro and P4-Phe side chains.
Shangen Xu; Junwei Zhou; Yingjin Chen; Xue Tong; Zixin Wang; Jiahui Guo; Jiyao Chen; Liurong Fang; Dang Wang; Shaobo Xiao. Characterization of Self-Processing Activities and Substrate Specificities of Porcine Torovirus 3C-Like Protease. Journal of Virology 2020, 94, 1 .
AMA StyleShangen Xu, Junwei Zhou, Yingjin Chen, Xue Tong, Zixin Wang, Jiahui Guo, Jiyao Chen, Liurong Fang, Dang Wang, Shaobo Xiao. Characterization of Self-Processing Activities and Substrate Specificities of Porcine Torovirus 3C-Like Protease. Journal of Virology. 2020; 94 (20):1.
Chicago/Turabian StyleShangen Xu; Junwei Zhou; Yingjin Chen; Xue Tong; Zixin Wang; Jiahui Guo; Jiyao Chen; Liurong Fang; Dang Wang; Shaobo Xiao. 2020. "Characterization of Self-Processing Activities and Substrate Specificities of Porcine Torovirus 3C-Like Protease." Journal of Virology 94, no. 20: 1.
Like obligate intracellular parasites, viruses co-opt host cell resources to establish productive infections. Polyamines are key aliphatic molecules that perform important roles in cellular growth and proliferation. They are also needed for the successful multiplication of various viruses. Little is known about the effects of polyamines on Arteriviridae infections. Here, porcine reproductive and respiratory syndrome virus (PRRSV), an economically prominent porcine virus, was used to investigate virus–polyamine interactions. We found that PRRSV infection significantly downregulated the levels of cellular polyamines. Using an inhibitor or specific short interfering RNAs (siRNAs) of ornithine decarboxylase 1, a key anabolic enzyme involved in the classical de novo biosynthesis of polyamines, we found that polyamine depletion abrogated PRRSV proliferation, and this effect was recoverable by adding exogenous spermidine and spermine, but not putrescine to the cells, suggesting that the host inhibits polyamine biosynthesis to restrict PRRSV proliferation. Further analysis revealed that the expression level of spermidine-spermine acetyltransferase 1 (SAT1), a catabolic enzyme that reduces spermidine and spermine levels, was upregulated during PRRSV infection, but conversely, SAT1 had an inhibitory effect on PRRSV reproduction. Our data show that polyamines are important molecules during PRRSV-host interactions, and polyamines and their biosynthetic pathways are potential therapeutic targets against PRRSV infection.
Yanrong Zhou; Zhenzhen Hou; Liurong Fang; Qiyun Ke; Yujian Xiong; Puxian Fang; Shaobo Xiao. Polyamine regulation of porcine reproductive and respiratory syndrome virus infection depends on spermidine-spermine acetyltransferase 1. Veterinary Microbiology 2020, 250, 108839 -108839.
AMA StyleYanrong Zhou, Zhenzhen Hou, Liurong Fang, Qiyun Ke, Yujian Xiong, Puxian Fang, Shaobo Xiao. Polyamine regulation of porcine reproductive and respiratory syndrome virus infection depends on spermidine-spermine acetyltransferase 1. Veterinary Microbiology. 2020; 250 ():108839-108839.
Chicago/Turabian StyleYanrong Zhou; Zhenzhen Hou; Liurong Fang; Qiyun Ke; Yujian Xiong; Puxian Fang; Shaobo Xiao. 2020. "Polyamine regulation of porcine reproductive and respiratory syndrome virus infection depends on spermidine-spermine acetyltransferase 1." Veterinary Microbiology 250, no. : 108839-108839.
Porcine reproductive and respiratory syndrome virus (PRRSV) and transmissible gastroenteritis virus (TGEV) are two highly infectious and lethal viruses causing major economic losses to pig production. Here, we report generation of double-gene-knockout (DKO) pigs harboring edited knockout alleles for known receptor proteins CD163 and pAPN and show that DKO pigs are completely resistant to genotype 2 PRRSV and TGEV. We found no differences in meat-production or reproductive-performance traits between wild-type and DKO pigs, but detected increased iron in DKO muscle. Additional infection challenge experiments showed that DKO pigs exhibited decreased susceptibility to porcine deltacoronavirus (PDCoV), thus offering unprecedented in vivo evidence of pAPN as one of PDCoV receptors. Beyond showing that multiple gene edits can be combined in a livestock animal to achieve simultaneous resistance to two major viruses, our study introduces a valuable model for investigating infection mechanisms of porcine pathogenic viruses that exploit pAPN or CD163 for entry.
Kui Xu; Yanrong Zhou; Yulian Mu; Zhiguo Liu; Shaohua Hou; Yujian Xiong; Liurong Fang; Changli Ge; Yinghui Wei; Xiuling Zhang; Changjiang Xu; Jingjing Che; Ziyao Fan; Guangming Xiang; Jiankang Guo; Haitao Shang; Hua Li; Shaobo Xiao; Julang Li; Kui Li. CD163 and pAPN double-knockout pigs are resistant to PRRSV and TGEV and exhibit decreased susceptibility to PDCoV while maintaining normal production performance. eLife 2020, 9, 1 .
AMA StyleKui Xu, Yanrong Zhou, Yulian Mu, Zhiguo Liu, Shaohua Hou, Yujian Xiong, Liurong Fang, Changli Ge, Yinghui Wei, Xiuling Zhang, Changjiang Xu, Jingjing Che, Ziyao Fan, Guangming Xiang, Jiankang Guo, Haitao Shang, Hua Li, Shaobo Xiao, Julang Li, Kui Li. CD163 and pAPN double-knockout pigs are resistant to PRRSV and TGEV and exhibit decreased susceptibility to PDCoV while maintaining normal production performance. eLife. 2020; 9 ():1.
Chicago/Turabian StyleKui Xu; Yanrong Zhou; Yulian Mu; Zhiguo Liu; Shaohua Hou; Yujian Xiong; Liurong Fang; Changli Ge; Yinghui Wei; Xiuling Zhang; Changjiang Xu; Jingjing Che; Ziyao Fan; Guangming Xiang; Jiankang Guo; Haitao Shang; Hua Li; Shaobo Xiao; Julang Li; Kui Li. 2020. "CD163 and pAPN double-knockout pigs are resistant to PRRSV and TGEV and exhibit decreased susceptibility to PDCoV while maintaining normal production performance." eLife 9, no. : 1.
During viral infection, the host cell synthesizes high amounts of viral proteins, which often causes stress to the endoplasmic reticulum (ER). To manage abnormal ER stress, mammalian cells trigger a response called the unfolded protein response (UPR). Previous studies have indicated that porcine reproductive and respiratory syndrome virus (PRRSV), an Arterivirus that has been devastating the swine industry worldwide, can induce ER stress and activate UPR, however, the activation pathways and the biological significance requires further investigation. In this study, we demonstrated that, among the three types of UPR pathways, PRRSV infection induced PERK and IRE1 pathways, but not the ATF6 pathway. Furthermore, the induction of UPR promoted PRRSV replication. We also found that PRRSV-induced UPR, particularly the PERK pathway, was involved in the induction of autophagy, a cellular degradation process that can alleviate cell stress. Besides, we also provided insights into the ER stress-mediated apoptosis in response to PRRSV infection. PRRSV infection induced the expression of the transcription factor CHOP, which activated caspase 3 and PARP led to ER stress-mediated apoptosis. Using 3-Methyladenine (3-MA) to inhibit autophagy, the increased ER stress and cell apoptosis were observed in the PRRSV infected cell. Taken together, our results revealed the associations of ER stress, autophagy, and apoptosis during PRRSV infection, helping us to further understand how PRRSV interacts with host cells.
Quangang Chen; Yanjuan Men; Dang Wang; Deqin Xu; Suyan Liu; Shaobo Xiao; Liurong Fang. Porcine reproductive and respiratory syndrome virus infection induces endoplasmic reticulum stress, facilitates virus replication, and contributes to autophagy and apoptosis. Scientific Reports 2020, 10, 1 -13.
AMA StyleQuangang Chen, Yanjuan Men, Dang Wang, Deqin Xu, Suyan Liu, Shaobo Xiao, Liurong Fang. Porcine reproductive and respiratory syndrome virus infection induces endoplasmic reticulum stress, facilitates virus replication, and contributes to autophagy and apoptosis. Scientific Reports. 2020; 10 (1):1-13.
Chicago/Turabian StyleQuangang Chen; Yanjuan Men; Dang Wang; Deqin Xu; Suyan Liu; Shaobo Xiao; Liurong Fang. 2020. "Porcine reproductive and respiratory syndrome virus infection induces endoplasmic reticulum stress, facilitates virus replication, and contributes to autophagy and apoptosis." Scientific Reports 10, no. 1: 1-13.
Non-structural protein 1 (nsp1) is only characterized in alphacoronaviruses (α-CoVs) and betacoronaviruses (β-CoVs). There have been extensive researches on how the β-CoVs nsp1 regulates viral virulence by inhibiting host protein synthesis, but the regulatory mechanism of the α-CoVs nsp1 is still unclear. Here, we report the 2.1-Å full-length crystal structure of nsp1 in emerging porcine SADS-CoV and the 1.8-Å full-length crystal structure of nsp1 in the highly lethal cat FIPV. Although they belong to different subtypes of α-CoVs, these viruses all have a bucket-shaped fold composed of six β-sheets, similar to the crystal structure of PEDV and TGEV nsp1. Comparing the above four structures, we found that the structure of α-CoVs nsp1 in the same subtype was more conserved. We then selected mammalian cells that were treated with SADS-CoV and FIPV nsp1 for RNA sequencing analysis and found that nsp1 had a specific inhibitory effect on interferon (IFN) and cell cycle genes. Using the Renilla luciferase (Rluc) assay and Western blotting, we confirmed that seven representative α-CoVs nsp1s could significantly inhibit the phosphorylation of STAT1-S727 and interfere with the effect of IFN-I. Moreover, the cell cycle experiment confirmed that α-CoVs nsp1 could encourage host cells to stay in the G0/G1 phase. Based on these findings, we not only greatly improved the crystal structure data on α-CoVs nsp1, but we also speculated that α-CoVs nsp1 regulated host proliferation and immune evasion-related biological functions by inhibiting the synthesis of host proteins, thus creating an environment conducive to the virus.
Zhou Shen; Yiling Yang; Siqi Yang; Guangxu Zhang; Shaobo Xiao; Zhen F. Fu; Guiqing Peng. Structural and Biological Basis of Alphacoronavirus nsp1 Associated with Host Proliferation and Immune Evasion. Viruses 2020, 12, 812 .
AMA StyleZhou Shen, Yiling Yang, Siqi Yang, Guangxu Zhang, Shaobo Xiao, Zhen F. Fu, Guiqing Peng. Structural and Biological Basis of Alphacoronavirus nsp1 Associated with Host Proliferation and Immune Evasion. Viruses. 2020; 12 (8):812.
Chicago/Turabian StyleZhou Shen; Yiling Yang; Siqi Yang; Guangxu Zhang; Shaobo Xiao; Zhen F. Fu; Guiqing Peng. 2020. "Structural and Biological Basis of Alphacoronavirus nsp1 Associated with Host Proliferation and Immune Evasion." Viruses 12, no. 8: 812.
Porcine reproductive and respiratory syndrome virus (PRRSV) is a major respiratory disease agent in pigs that causes enormous economic losses to the global swine industry. PRRSV helicase nsp10 is a multifunctional protein with translocation and unwinding activities and plays a vital role in viral RNA synthesis. Here, we report the first structure of full-length nsp10 from the arterivirus PRRSV at 3.0-Å resolution. Our results show that the 1B domain of PRRSV nsp10 adopts a novel open state and has a unique C-terminal domain structure, which plays a crucial role in nsp10 helicase activity. Furthermore, mutagenesis and structural analysis revealed conservation of the helicase catalytic domain across the order Nidovirales (families Arteriviridae and Coronaviridae ). Importantly, our results will provide a structural basis for further understanding the function of helicases in the order Nidovirales .
Yuejun Shi; Xiaohan Tong; Gang Ye; Ruixue Xiu; Lisha Li; Limeng Sun; Jiale Shi; Mengxia Li; Yunfeng Song; Chengpeng Fan; Ke Shi; Zhen F. Fu; Shaobo Xiao; Guiqing Peng. Structural Characterization of the Helicase nsp10 Encoded by Porcine Reproductive and Respiratory Syndrome Virus. Journal of Virology 2020, 94, 1 .
AMA StyleYuejun Shi, Xiaohan Tong, Gang Ye, Ruixue Xiu, Lisha Li, Limeng Sun, Jiale Shi, Mengxia Li, Yunfeng Song, Chengpeng Fan, Ke Shi, Zhen F. Fu, Shaobo Xiao, Guiqing Peng. Structural Characterization of the Helicase nsp10 Encoded by Porcine Reproductive and Respiratory Syndrome Virus. Journal of Virology. 2020; 94 (15):1.
Chicago/Turabian StyleYuejun Shi; Xiaohan Tong; Gang Ye; Ruixue Xiu; Lisha Li; Limeng Sun; Jiale Shi; Mengxia Li; Yunfeng Song; Chengpeng Fan; Ke Shi; Zhen F. Fu; Shaobo Xiao; Guiqing Peng. 2020. "Structural Characterization of the Helicase nsp10 Encoded by Porcine Reproductive and Respiratory Syndrome Virus." Journal of Virology 94, no. 15: 1.
Interferon (IFN)-stimulated gene (ISG) induction through IFN signaling is important to create an antiviral state and usually directly inhibits virus infection. The present study first demonstrated that PDCoV nsp5 can cleave mRNA-decapping enzyme 1a (DCP1A) to attenuate its antiviral activity. Furthermore, cleaving DCP1A is a common characteristic of nsp5 proteins from different coronaviruses (CoVs), which represents a common immune evasion mechanism of CoVs. Previous evidence showed that CoV nsp5 cleaves the NF-κB essential modulator and signal transducer and activator of transcription 2. Taken together, CoV nsp5 is a potent IFN antagonist because it can simultaneously target different aspects of the host IFN system, including IFN production and signaling and effector molecules.
Xinyu Zhu; Jiyao Chen; Liyuan Tian; Yanrong Zhou; Shangen Xu; Siwen Long; Dang Wang; Liurong Fang; Shaobo Xiao. Porcine Deltacoronavirus nsp5 Cleaves DCP1A To Decrease Its Antiviral Activity. Journal of Virology 2020, 94, 1 .
AMA StyleXinyu Zhu, Jiyao Chen, Liyuan Tian, Yanrong Zhou, Shangen Xu, Siwen Long, Dang Wang, Liurong Fang, Shaobo Xiao. Porcine Deltacoronavirus nsp5 Cleaves DCP1A To Decrease Its Antiviral Activity. Journal of Virology. 2020; 94 (15):1.
Chicago/Turabian StyleXinyu Zhu; Jiyao Chen; Liyuan Tian; Yanrong Zhou; Shangen Xu; Siwen Long; Dang Wang; Liurong Fang; Shaobo Xiao. 2020. "Porcine Deltacoronavirus nsp5 Cleaves DCP1A To Decrease Its Antiviral Activity." Journal of Virology 94, no. 15: 1.
Porcine deltacoronavirus (PDCoV) is a novel swine enteropathogenic coronavirus that causes watery diarrhea, vomiting and mortality in nursing piglets. Type III interferons (IFN-λs) are the major antiviral cytokines in intestinal epithelial cells, the target cells in vivo for PDCoV. In this study, we found that PDCoV infection remarkably inhibited Sendai virus-induced IFN-λ1 production by suppressing transcription factors IRF and NF-κB in IPI-2I cells, a line of porcine intestinal mucosal epithelial cells. We also confirmed that PDCoV infection impeded the activation of IFN-λ1 promoter stimulated by RIG-I, MDA5 and MAVS, but not by TBK1 and IRF1. Although the expression levels of IRF1 and MAVS were not changed, PDCoV infection resulted in reduction of the number of peroxisomes, the platform for MAVS to activate IRF1, and subsequent type III IFN production. Taken together, our study demonstrates that PDCoV suppresses type III IFN responses to circumvent the host’s antiviral immunity.
Shudan Liu; Puxian Fang; Wenting Ke; Jing Wang; Xunlei Wang; Shaobo Xiao; Liurong Fang. Porcine deltacoronavirus (PDCoV) infection antagonizes interferon-λ1 production. Veterinary Microbiology 2020, 247, 108785 -108785.
AMA StyleShudan Liu, Puxian Fang, Wenting Ke, Jing Wang, Xunlei Wang, Shaobo Xiao, Liurong Fang. Porcine deltacoronavirus (PDCoV) infection antagonizes interferon-λ1 production. Veterinary Microbiology. 2020; 247 ():108785-108785.
Chicago/Turabian StyleShudan Liu; Puxian Fang; Wenting Ke; Jing Wang; Xunlei Wang; Shaobo Xiao; Liurong Fang. 2020. "Porcine deltacoronavirus (PDCoV) infection antagonizes interferon-λ1 production." Veterinary Microbiology 247, no. : 108785-108785.
Despite the good biocompatibility and antibacterial activity of zinc sulfide nanoparticles (ZnS NPs), whether they possess antiviral activity is still unclear. Here, GSH-modified ZnS NPs (GSH-ZnS NPs) were synthesized and their significant antiviral activity was demonstrated using the Arteriviridae family RNA virus, porcine reproductive and respiratory syndrome virus (PRRSV), as a model. Mechanistically, GSH-ZnS NPs were shown to reduce PRRSV-induced ROS production to prevent PRRSV multiplication, with no activating effect on the interferon (IFN) signal pathway, the first defense line against virus infection. Furthermore, isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomic analysis of GSH-ZnS NP-treated cells revealed the involvement of numerous crucial proteins in virus proliferation, with vitronectin (VTN) being confirmed as an efficient PRRSV antagonist here. Furthermore, GSH-ZnS NPs were found to have potent antiviral effects on the Herpesviridae family DNA virus, pseudorabies virus (PRV), the Coronaviridae family positive-sense RNA virus, porcine epidemic diarrhea virus (PEDV), and the Rhabdoviridae family negative-stranded RNA virus, vesicular stomatitis virus (VSV), indicating their broad-spectrum antiviral activity against viruses from different families with various genome types. Overall, GSH-ZnS NP is a prospective candidate for the development of antiviral nanomaterials and may serve as a model for investigation of potential host restriction factors in combination with proteomics.
Yanrong Zhou; Ting Tong; Xiaohan Jiang; Liurong Fang; Yuan Wu; Jiangong Liang; Shaobo Xiao. GSH-ZnS Nanoparticles Exhibit High-Efficiency and Broad-Spectrum Antiviral Activities via Multistep Inhibition Mechanisms. ACS Applied Bio Materials 2020, 3, 4809 -4819.
AMA StyleYanrong Zhou, Ting Tong, Xiaohan Jiang, Liurong Fang, Yuan Wu, Jiangong Liang, Shaobo Xiao. GSH-ZnS Nanoparticles Exhibit High-Efficiency and Broad-Spectrum Antiviral Activities via Multistep Inhibition Mechanisms. ACS Applied Bio Materials. 2020; 3 (8):4809-4819.
Chicago/Turabian StyleYanrong Zhou; Ting Tong; Xiaohan Jiang; Liurong Fang; Yuan Wu; Jiangong Liang; Shaobo Xiao. 2020. "GSH-ZnS Nanoparticles Exhibit High-Efficiency and Broad-Spectrum Antiviral Activities via Multistep Inhibition Mechanisms." ACS Applied Bio Materials 3, no. 8: 4809-4819.
As an emerging swine enteropathogenic coronavirus, porcine deltacoronavirus (PDCoV) not only causes serious diarrhea in suckling piglets but also possesses the potential for cross-species transmission, which has sparked growing interest when studying this emerging virus. We previously identified a novel accessory protein NS7a encoded by PDCoV; however, the function of NS7a was not resolved. In this study, we demonstrated that PDCoV NS7a is an interferon antagonist. Overexpression of NS7a notably inhibited Sendai virus (SeV)-induced interferon-β (IFN-β) production and the activation of IRF3 rather than NF-κB. NS7a also inhibited IFN-β promoter activity induced by RIG-I, MDA5, MAVS, TBK1, and IKKε, which are key components of the RIG-I-like receptor (RLR) signaling pathway but not IRF3, the transcription factor downstream of TBK1/IKKε. Surprisingly, NS7a specifically interacts with IKKε but not with the closely related TBK1. Furthermore, NS7a interacts simultaneously with the kinase domain (KD) and the scaffold dimerization domain (SDD) of IKKε, competing with TRAF3, and IRF3 for binding to IKKε, leading to the reduction of RLR-mediated IFN-β production. The interactions of TRAF3-IKKε and IKKε-IRF3 are also attenuated in PDCoV-infected cells. Taken together, our results demonstrate that PDCoV NS7a inhibits IFN-β production by disrupting the association of IKKε with both TRAF3 and IRF3, revealing a new mechanism utilized by a PDCoV accessory protein to evade the host antiviral innate immune response.
Puxian Fang; Liurong Fang; Sijin Xia; Jie Ren; Jiansong Zhang; Dongcheng Bai; Yanrong Zhou; Guiqing Peng; Shuhong Zhao; Shaobo Xiao. Porcine Deltacoronavirus Accessory Protein NS7a Antagonizes IFN-β Production by Competing With TRAF3 and IRF3 for Binding to IKKε. Frontiers in Cellular and Infection Microbiology 2020, 10, 257 .
AMA StylePuxian Fang, Liurong Fang, Sijin Xia, Jie Ren, Jiansong Zhang, Dongcheng Bai, Yanrong Zhou, Guiqing Peng, Shuhong Zhao, Shaobo Xiao. Porcine Deltacoronavirus Accessory Protein NS7a Antagonizes IFN-β Production by Competing With TRAF3 and IRF3 for Binding to IKKε. Frontiers in Cellular and Infection Microbiology. 2020; 10 ():257.
Chicago/Turabian StylePuxian Fang; Liurong Fang; Sijin Xia; Jie Ren; Jiansong Zhang; Dongcheng Bai; Yanrong Zhou; Guiqing Peng; Shuhong Zhao; Shaobo Xiao. 2020. "Porcine Deltacoronavirus Accessory Protein NS7a Antagonizes IFN-β Production by Competing With TRAF3 and IRF3 for Binding to IKKε." Frontiers in Cellular and Infection Microbiology 10, no. : 257.