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I have a broad background in virology and cell biology, with specific training and expertise in HIV-host interactions. My current research is focusing on the mechanism of host restrictions on viral replication. As PI or co-Investigator on several university- and NIH-funded grants, we have uncovered the anti-HIV-1 activities of human APOBEC3F, APOBEC3DE, and APOBEC3H. We also discovered the antiretroviral activity of MOV10, which is a super-family I RNA helicase and a component of the RNA-induced silencing complex (RISC). Recently, my research moved to understand how HIV-1 Env glycoproteins are expressed from the secretory pathway and how they are inhibited by the host ER-associated protein degradation (ERAD) pathway. More recently, my lab returned back to the Nef field to understand how Nef counteracts the SERINC5 antiviral activity and how SERINC5 iinhibits HIV-1 entry.
Summary HIV-1-negative factor (Nef) protein antagonizes serine incorporator 5 (SERINC5) by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identify cyclin K (CycK) and cyclin-dependent kinase 13 (CDK13) as a Nef-associated kinase complex. CycK/CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counteractivity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we generate chimeric proteins between CD8 and SERINC5 to study their response to Nef. Nef not only downregulates but, importantly, also binds to this chimera in an S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.
Qingqing Chai; Sunan Li; Morgan K. Collins; Rongrong Li; Iqbal Ahmad; Silas F. Johnson; Dylan A. Frabutt; Zhichang Yang; Xiaojing Shen; Liangliang Sun; Jian Hu; Judd F. Hultquist; B. Matija Peterlin; Yong-Hui Zheng. HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity. Cell Reports 2021, 36, 1 .
AMA StyleQingqing Chai, Sunan Li, Morgan K. Collins, Rongrong Li, Iqbal Ahmad, Silas F. Johnson, Dylan A. Frabutt, Zhichang Yang, Xiaojing Shen, Liangliang Sun, Jian Hu, Judd F. Hultquist, B. Matija Peterlin, Yong-Hui Zheng. HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity. Cell Reports. 2021; 36 (6):1.
Chicago/Turabian StyleQingqing Chai; Sunan Li; Morgan K. Collins; Rongrong Li; Iqbal Ahmad; Silas F. Johnson; Dylan A. Frabutt; Zhichang Yang; Xiaojing Shen; Liangliang Sun; Jian Hu; Judd F. Hultquist; B. Matija Peterlin; Yong-Hui Zheng. 2021. "HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity." Cell Reports 36, no. 6: 1.
Summary HIV-1 Nef antagonizes SERINC5 by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identified cyclin K and cyclin-dependent kinase 13 (CycK:CDK13) as a new Nef-associated kinase complex. CycK:CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counter activity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we created chimeric proteins between CD8 and SERINC5. Nef not only downregulates, but importantly, also binds to this chimera in a S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.
Qingqing Chai; Sunan Li; Morgan K. Collins; Rongrong Li; Iqbal Ahmad; Silas F. Johnson; Dylan A. Frabutt; Zhichang Yang; Xiaojing Shen; Liangliang Sun; Jian Hu; Judd F. Hultquist; B. Matija Peterlin; Yong-Hui Zheng. HIV-1 Nef and CycK:CDK13 antagonize SERINC5 for optimal viral infectivity. 2021, 1 .
AMA StyleQingqing Chai, Sunan Li, Morgan K. Collins, Rongrong Li, Iqbal Ahmad, Silas F. Johnson, Dylan A. Frabutt, Zhichang Yang, Xiaojing Shen, Liangliang Sun, Jian Hu, Judd F. Hultquist, B. Matija Peterlin, Yong-Hui Zheng. HIV-1 Nef and CycK:CDK13 antagonize SERINC5 for optimal viral infectivity. . 2021; ():1.
Chicago/Turabian StyleQingqing Chai; Sunan Li; Morgan K. Collins; Rongrong Li; Iqbal Ahmad; Silas F. Johnson; Dylan A. Frabutt; Zhichang Yang; Xiaojing Shen; Liangliang Sun; Jian Hu; Judd F. Hultquist; B. Matija Peterlin; Yong-Hui Zheng. 2021. "HIV-1 Nef and CycK:CDK13 antagonize SERINC5 for optimal viral infectivity." , no. : 1.
Xin Yin; Zhe Hu; Qinyong Gu; Xingliang Wu; Yong-Hui Zheng; Ping Wei; Xiaojun Wang. Correction for Yin et al., “Equine Tetherin Blocks Retrovirus Release and Its Activity Is Antagonized by Equine Infectious Anemia Virus Envelope Protein”. Journal of Virology 2021, 95, 1 .
AMA StyleXin Yin, Zhe Hu, Qinyong Gu, Xingliang Wu, Yong-Hui Zheng, Ping Wei, Xiaojun Wang. Correction for Yin et al., “Equine Tetherin Blocks Retrovirus Release and Its Activity Is Antagonized by Equine Infectious Anemia Virus Envelope Protein”. Journal of Virology. 2021; 95 (6):1.
Chicago/Turabian StyleXin Yin; Zhe Hu; Qinyong Gu; Xingliang Wu; Yong-Hui Zheng; Ping Wei; Xiaojun Wang. 2021. "Correction for Yin et al., “Equine Tetherin Blocks Retrovirus Release and Its Activity Is Antagonized by Equine Infectious Anemia Virus Envelope Protein”." Journal of Virology 95, no. 6: 1.
The serine incorporator (SERINC) protein family has five paralogous members with 9–11 transmembrane domains. SERINC5 is a potent host restriction factor and antagonized by HIV-1 Nef and two other retroviral accessory proteins via the lysosomal degradation pathway. Here, we investigated human SERINC4 expression and antiviral mechanisms. Unlike its four paralogs, human SERINC4 is subjected to proteasome-mediated turnover, resulting in ~250-fold lower expression than SERINC5. However, when expression was normalized, human SERINC4 restricted HIV-1 replication as effectively as SERINC5, and SERINC4 was also antagonized by Nef via the lysosomal pathway. Although SERINC4 proteins are conserved within primates or rodents, their N-terminal regions are highly variable across species. Interestingly, unlike human SERINC4, murine SERINC4 was stably expressed but had a very poor antiviral activity. We created stable SERINC4 chimeras by replacing the N-terminal region and found that the 1–34 and 35–92 amino acids determine SERINC4 antiviral activity or protein expression, respectively. Using these chimeras, we demonstrate that SERINC4 is incorporated into HIV-1 virions and restricts Tier 1 HIV-1 more effectively than Tier 3 HIV-1. Importantly, SERINC4 increases HIV-1 sensitivity to broadly neutralizing antibodies. Thus, human SERINC4 strongly restricts HIV-1 replication when it is overexpressed, which reflects a potential antiviral activity of this gene product under physiological conditions.
Xusheng Qiu; Ifeanyichukwu E. Eke; Silas F. Johnson; Chan Ding; Yong-Hui Zheng. Proteasomal degradation of human SERINC4: A potent host anti-HIV-1 factor that is antagonized by nef. Current Research in Virological Science 2020, 1, 100002 .
AMA StyleXusheng Qiu, Ifeanyichukwu E. Eke, Silas F. Johnson, Chan Ding, Yong-Hui Zheng. Proteasomal degradation of human SERINC4: A potent host anti-HIV-1 factor that is antagonized by nef. Current Research in Virological Science. 2020; 1 ():100002.
Chicago/Turabian StyleXusheng Qiu; Ifeanyichukwu E. Eke; Silas F. Johnson; Chan Ding; Yong-Hui Zheng. 2020. "Proteasomal degradation of human SERINC4: A potent host anti-HIV-1 factor that is antagonized by nef." Current Research in Virological Science 1, no. : 100002.
The serine incorporator (SERINC) protein family has five paralogous members with 9-11 transmembrane domains. SERINC5 is a potent host restriction factor and antagonized by HIV-1 Nef and two other retroviral accessory proteins via the lysosomal degradation pathway. Here, we investigated human SERINC4 expression and antiviral mechanisms. Unlike its four paralogs, human SERINC4 is subjected to proteasome-mediated turnover, resulting in ~250-fold lower expression than SERINC5. However, when expression was normalized, human SERINC4 restricted HIV-1 replication as effectively as SERINC5, and SERINC4 was also antagonized by Nef via the lysosomal pathway. Although SERINC4 proteins are conserved within primates or rodents, their N-terminal regions are highly variable across species. Interestingly, unlike human SERINC4, murine SERINC4 was stably expressed but had a very poor antiviral activity. We created stable SERINC4 chimeras by replacing the N-terminal region and found that the 1-34 and 35-92 amino acids determine SERINC4 antiviral activity or protein expression, respectively. Using these chimeras, we demonstrate that SERINC4 is incorporated into HIV-1 virions and restricts Tier 1 HIV-1 more effectively than Tier 3 HIV-1. Importantly, SERINC4 increases HIV-1 sensitivity to broadly neutralizing antibodies. Thus, human SERINC4 strongly restricts HIV-1 replication when it is overexpressed, which reflects a potential antiviral activity of this gene product under physiological conditions.
Xusheng Qiu; Ifeanyichukwu E. Eke; Silas F. Johnson; Chan Ding; Yong-Hui Zheng. Proteasomal degradation of human SERINC4: a potent host anti-HIV-1 factor that is antagonized by Nef. 2020, 1 .
AMA StyleXusheng Qiu, Ifeanyichukwu E. Eke, Silas F. Johnson, Chan Ding, Yong-Hui Zheng. Proteasomal degradation of human SERINC4: a potent host anti-HIV-1 factor that is antagonized by Nef. . 2020; ():1.
Chicago/Turabian StyleXusheng Qiu; Ifeanyichukwu E. Eke; Silas F. Johnson; Chan Ding; Yong-Hui Zheng. 2020. "Proteasomal degradation of human SERINC4: a potent host anti-HIV-1 factor that is antagonized by Nef." , no. : 1.
Enveloped viruses express three classes of fusion proteins that are required for their entry into host cells via mediating virus and cell membrane fusion. Class I fusion proteins are produced from influenza viruses, retroviruses, Ebola viruses, and coronaviruses. They are first synthesized as a type I transmembrane polypeptide precursor that is subsequently glycosylated and oligomerized. Most of these precursors are cleaved en route to the plasma membrane by a cellular protease furin in the late secretory pathway, generating the trimeric N-terminal receptor-binding and C-terminal fusion subunits. Here, we show that a cellular protein, MARCH8, specifically inhibits the furin-mediated cleavage of EBOV GP, HIV-1 Env, and H5N1 HA. Further analyses uncovered that MARCH8 blocked the EBOV GP glycosylation in the Golgi and inhibited its transport from the Golgi to the plasma membrane. Thus, MARCH8 has a very broad antiviral activity by specifically inactivating different viral fusion proteins.
Changqing Yu; Sunan Li; Xianfeng Zhang; İlyas Khan; Iqbal Ahmad; Yulong Zhou; Shuo Li; Jing Shi; Yu Wang; Yong-Hui Zheng. MARCH8 Inhibits Ebola Virus Glycoprotein, Human Immunodeficiency Virus Type 1 Envelope Glycoprotein, and Avian Influenza Virus H5N1 Hemagglutinin Maturation. mBio 2020, 11, 1 .
AMA StyleChangqing Yu, Sunan Li, Xianfeng Zhang, İlyas Khan, Iqbal Ahmad, Yulong Zhou, Shuo Li, Jing Shi, Yu Wang, Yong-Hui Zheng. MARCH8 Inhibits Ebola Virus Glycoprotein, Human Immunodeficiency Virus Type 1 Envelope Glycoprotein, and Avian Influenza Virus H5N1 Hemagglutinin Maturation. mBio. 2020; 11 (5):1.
Chicago/Turabian StyleChangqing Yu; Sunan Li; Xianfeng Zhang; İlyas Khan; Iqbal Ahmad; Yulong Zhou; Shuo Li; Jing Shi; Yu Wang; Yong-Hui Zheng. 2020. "MARCH8 Inhibits Ebola Virus Glycoprotein, Human Immunodeficiency Virus Type 1 Envelope Glycoprotein, and Avian Influenza Virus H5N1 Hemagglutinin Maturation." mBio 11, no. 5: 1.
The transmission and evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are of paramount importance in controlling and combating the coronavirus disease 2019 (COVID-19) pandemic. Currently, over 15,000 SARS-CoV-2 single mutations have been recorded, which have a great impact on the development of diagnostics, vaccines, antibody therapies, and drugs. However, little is known about SARS-CoV-2’s evolutionary characteristics and general trend. In this work, we present a comprehensive genotyping analysis of existing SARS-CoV-2 mutations. We reveal that host immune response via APOBEC and ADAR gene editing gives rise to near 65% of recorded mutations. Additionally, we show that children under age five and the elderly may be at high risk from COVID-19 because of their overreaction to the viral infection. Moreover, we uncover that populations of Oceania and Africa react significantly more intensively to SARS-CoV-2 infection than those of Europe and Asia, which may explain why African Americans were shown to be at increased risk of dying from COVID-19, in addition to their high risk of COVID-19 infection caused by systemic health and social inequities. Finally, our study indicates that for two viral genome sequences of the same origin, their evolution order may be determined from the ratio of mutation type, C > T over T > C.
Rui Wang; Yuta Hozumi; Yong-Hui Zheng; Changchuan Yin; Guo-Wei Wei. Host Immune Response Driving SARS-CoV-2 Evolution. Viruses 2020, 12, 1095 .
AMA StyleRui Wang, Yuta Hozumi, Yong-Hui Zheng, Changchuan Yin, Guo-Wei Wei. Host Immune Response Driving SARS-CoV-2 Evolution. Viruses. 2020; 12 (10):1095.
Chicago/Turabian StyleRui Wang; Yuta Hozumi; Yong-Hui Zheng; Changchuan Yin; Guo-Wei Wei. 2020. "Host Immune Response Driving SARS-CoV-2 Evolution." Viruses 12, no. 10: 1095.
Ebola virus (EBOV) glycoprotein (GP) is a class I fusion protein whose maturation is dependent on furin-mediated processing. EBOV-GP is heavily glycosylated, with glycans constituting ~50% of its molecular mass. Compared with 15 N-linked glycosylation sites, EBOV-GP1 has ~80 potential O-linked glycosylation sites in the mucin-like domain (MLD), suggesting that O-linked glycans are dominated. The membrane-associated RING-CH (MARCH) family consists of 11 members that are RING-finger ubiquitin E3 ligases. Recently, human MARCH1, MARCH2, and MARCH8 were reported to inhibit HIV-1 replication by targeting its Env. Here, we show that human MARCH8 also inhibits EBOV replication by blocking GP incorporation into virions via downregulating its cell surface expression. To understand how the downregulation occurs, we investigated EBOV-GP subcellular localization, processing, glycosylation, and intracellular trafficking in the presence of human MARCH8. We find that MARCH8 interacts with GP and retains GP in the Golgi. MARCH8 also interacts with the homoB domain of furin that blocks its convertase activity. In consequence, MARCH8 blocks GP processing in an MLD-independent manner. Consistently, MARCH8 also blocks the O-linked, but not the N-linked glycosylation of GP. Importantly, in the presence of MARCH8, the shedding of GP1 but not the secreted GP (sGP) is blocked, suggesting that MARCH8 targets the GP1 C-terminal region. The MARCH8 activity is extended to its orthologs from Bos taurus and mice, and its paralogs MARCH1 and MARCH2. In addition, MARCH8 inhibits the processing of two other class I fusion proteins, including HIV-1 Env and IAV HA, and it triggers the degradation of the class III fusion protein VSV-G. We conclude that MARCH8 exerts a very broad and conserved antiviral activity by inhibiting the maturation of class I fusion proteins, which blocks their secretion to the cell surface and incorporation into virions. It should also target class III fusion proteins by triggering their degradation.
Changqing Yu; Sunan Li; Omid Madadgar; Iqbal Ahmad; Xianfeng Zhang; Jing Shi; Yu Wang; Yulong Zhou; Yong-Hui Zheng. MARCH8 Restricts Ebola Virus Replication by Blocking the Viral Glycoprotein Processing and Glycosylation. Proceedings 2020, 50, 123 .
AMA StyleChangqing Yu, Sunan Li, Omid Madadgar, Iqbal Ahmad, Xianfeng Zhang, Jing Shi, Yu Wang, Yulong Zhou, Yong-Hui Zheng. MARCH8 Restricts Ebola Virus Replication by Blocking the Viral Glycoprotein Processing and Glycosylation. Proceedings. 2020; 50 (1):123.
Chicago/Turabian StyleChangqing Yu; Sunan Li; Omid Madadgar; Iqbal Ahmad; Xianfeng Zhang; Jing Shi; Yu Wang; Yulong Zhou; Yong-Hui Zheng. 2020. "MARCH8 Restricts Ebola Virus Replication by Blocking the Viral Glycoprotein Processing and Glycosylation." Proceedings 50, no. 1: 123.
The nuclear export receptor CRM1 is an important regulator involved in the shuttling of various cellular and viral RNAs between the nucleus and the cytoplasm. HIV-1 Rev interacts with CRM1 in the late phase of HIV-1 replication to promote nuclear export of unspliced and single spliced HIV-1 transcripts. However, other cellular factors involved in the CRM1-dependent viral RNA nuclear export remain largely unknown. Here, we demonstrate that ANP32A and ANP32B mediate the export of unspliced or partially spliced viral mRNA via interactions with Rev and CRM1. We found that a double, but not single, knockout of ANP32A and ANP32B significantly decreased the expression of gag protein. Reconstitution of either ANP32A or ANP32B restored the viral production equally. Disruption of both ANP32A and ANP32B expression led to a dramatic accumulation of unspliced viral mRNA in the nucleus. We further identified that ANP32A and ANP32B interact with both Rev and CRM1 to promote RNA transport. Our data strongly suggest that ANP32A and ANP32B play an important role in the Rev–CRM1 pathway, which is essential for HIV-1 replication, and our findings provide a candidate therapeutic target for host defense against retroviral infection.
Yujie Wang; Haili Zhang; Lei Na; Cheng Du; Zhenyu Zhang; Yong-Hui Zheng; Xiaojun Wang. ANP32A and ANP32B are key factors in the Rev-dependent CRM1 pathway for nuclear export of HIV-1 unspliced mRNA. Journal of Biological Chemistry 2019, 294, 15346 -15357.
AMA StyleYujie Wang, Haili Zhang, Lei Na, Cheng Du, Zhenyu Zhang, Yong-Hui Zheng, Xiaojun Wang. ANP32A and ANP32B are key factors in the Rev-dependent CRM1 pathway for nuclear export of HIV-1 unspliced mRNA. Journal of Biological Chemistry. 2019; 294 (42):15346-15357.
Chicago/Turabian StyleYujie Wang; Haili Zhang; Lei Na; Cheng Du; Zhenyu Zhang; Yong-Hui Zheng; Xiaojun Wang. 2019. "ANP32A and ANP32B are key factors in the Rev-dependent CRM1 pathway for nuclear export of HIV-1 unspliced mRNA." Journal of Biological Chemistry 294, no. 42: 15346-15357.
Restriction factors provide the first line of defense against retrovirus infection by posing several blocks to the viral replication cycle. SERINC5 is a novel restriction factor that strongly blocks HIV-1 entry, although it is counteracted by Nef. Currently, it is still unclear how HIV-1 entry is blocked by SERINC5. Notably, this entry block is dependent on viral Env proteins. Laboratory-adapted HIV-1 strains are sensitive, whereas primary isolates are highly resistant to SERINC5. Env proteins mediate virus entry via extensive conformational rearrangements from a closed ground state to a CD4-bound open state. We detected Env-Env associations and Env-SERINC5 interactions in live cells by a novel bimolecular fluorescence assay. We demonstrate that CD4 expression increases the Env sensitivity to SERINC5 and allows SERINC5 to dissociate the Env complex, suggesting that SERINC5 restriction is dependent on Env conformation. Our results provide new insights into the poorly defined Env-dependent SERINC5 antiviral mechanism.
Xihe Zhang; Jing Shi; Xusheng Qiu; Qingqing Chai; Dylan Frabutt; Richard C. Schwartz; Yong-Hui Zheng. CD4 Expression and Env Conformation Are Critical for HIV-1 Restriction by SERINC5. Journal of Virology 2019, 93, 1 .
AMA StyleXihe Zhang, Jing Shi, Xusheng Qiu, Qingqing Chai, Dylan Frabutt, Richard C. Schwartz, Yong-Hui Zheng. CD4 Expression and Env Conformation Are Critical for HIV-1 Restriction by SERINC5. Journal of Virology. 2019; 93 (14):1.
Chicago/Turabian StyleXihe Zhang; Jing Shi; Xusheng Qiu; Qingqing Chai; Dylan Frabutt; Richard C. Schwartz; Yong-Hui Zheng. 2019. "CD4 Expression and Env Conformation Are Critical for HIV-1 Restriction by SERINC5." Journal of Virology 93, no. 14: 1.
Bovine leukemia virus (BLV) causes enzootic bovine leukosis in cattle and leads to heavy economic losses in the husbandry industry. Heilongjiang Province, China, is rich in dairy cattle. However, its current BLV epidemiology and genotypes have still not been evaluated and confirmed. In this report, we investigated the BLV epidemiology in dairy cattle in the major regions of Heilongjiang Province via the nested PCR assay. A total of 730 blood samples were collected from nine different farms in six regions of Heilongjiang. The results showed that the infection rate of these regions ranged from null to 31%. With a clustering analysis of 60 published BLV env sequences, genotypes 1 and 6 were confirmed to be circulating in Heilongjiang. Importantly, a new genotype, 11, and a new subgenotype, 6E, were also identified in the Harbin and Daqing regions, respectively. An epitope analysis showed that a cluster of T-X-D-X-R-XXXX-A sequences in genotype 11 gp51 neutralizing domain 2 was unique among all currently known BLV isolates and was therefore a defining feature of this new genotype. BLV epidemics and genotypes were initially investigated in dairy cattle of Heilongjiang. A relatively high infection rate was found in some regions of this province. A new genotype, G11, with a highly specific motif, was identified and thus added as a new member to the current BLV genotype family. This report provides an initial reference for future investigations and subsequent control of BLV transmission and spread in this region.
Changqing Yu; Xuefeng Wang; Yulong Zhou; Yu Wang; Xianfeng Zhang; Yonghui Zheng. Genotyping bovine leukemia virus in dairy cattle of Heilongjiang, northeastern China. BMC Veterinary Research 2019, 15, 179 .
AMA StyleChangqing Yu, Xuefeng Wang, Yulong Zhou, Yu Wang, Xianfeng Zhang, Yonghui Zheng. Genotyping bovine leukemia virus in dairy cattle of Heilongjiang, northeastern China. BMC Veterinary Research. 2019; 15 (1):179.
Chicago/Turabian StyleChangqing Yu; Xuefeng Wang; Yulong Zhou; Yu Wang; Xianfeng Zhang; Yonghui Zheng. 2019. "Genotyping bovine leukemia virus in dairy cattle of Heilongjiang, northeastern China." BMC Veterinary Research 15, no. 1: 179.
Serine incorporator 5 (SERINC5) is a recently identified restriction factor that blocks virus entry but is antagonized by three unrelated retroviral accessory proteins. The S2 protein from equine infectious anemia virus (EIAV) has been reported to reduce SERINC5 expression at steady-state levels likely via the endocytic pathway; however, the precise mechanism is still unclear. Here, we investigated how EIAV S2 protein down-regulates SERINC5 compared with down-regulation induced by Nef from HIV-1 and glycoMA proteins from murine leukemia virus (MLV). Using bimolecular fluorescence complementation (BiFC) assay and immunoprecipitation (IP), we detected an interaction between S2 and SERINC5. We found that this interaction relies on the S2 myristoylation site, indicating that it may occur on the plasma membrane. S2 internalized SERINC5 via receptor-mediated endocytosis and targeted it to endosomes and lysosomes, resulting in a ubiquitination-dependent decrease in SERINC5 expression at steady-state levels. Both BiFC and IP detected a glycoMA–SERINC5 interaction, but a Nef–SERINC5 interaction was detected only by BiFC. Moreover, S2 and glycoMA down-regulated SERINC5 more effectively than did Nef. We further show that unlike Nef, both S2 and glycoMA effectively down-regulate SERINC2 and also SERINC5 from Xenopus tropicalis (xSERINC5). Moreover, we detected expression of the equine SERINC5 (eSERINC5) protein and observed that its expression is much weaker than expression levels of SERINC5 from other species. Nonetheless, eSERINC5 had a strong antiviral activity that was effectively counteracted by S2. We conclude that HIV-1, EIAV, and MLV share a similar mechanism to antagonize viral restriction by host SERINC5.
Iqbal Ahmad; Sunan Li; Rongrong Li; Qingqing Chai; Lixin Zhang; Bin Wang; Changqing Yu; Yong-Hui Zheng. The retroviral accessory proteins S2, Nef, and glycoMA use similar mechanisms for antagonizing the host restriction factor SERINC5. Journal of Biological Chemistry 2019, 294, 7013 -7024.
AMA StyleIqbal Ahmad, Sunan Li, Rongrong Li, Qingqing Chai, Lixin Zhang, Bin Wang, Changqing Yu, Yong-Hui Zheng. The retroviral accessory proteins S2, Nef, and glycoMA use similar mechanisms for antagonizing the host restriction factor SERINC5. Journal of Biological Chemistry. 2019; 294 (17):7013-7024.
Chicago/Turabian StyleIqbal Ahmad; Sunan Li; Rongrong Li; Qingqing Chai; Lixin Zhang; Bin Wang; Changqing Yu; Yong-Hui Zheng. 2019. "The retroviral accessory proteins S2, Nef, and glycoMA use similar mechanisms for antagonizing the host restriction factor SERINC5." Journal of Biological Chemistry 294, no. 17: 7013-7024.
The nuclear export receptor CRM1 is an important regulator involved in the shuttling of various cellular and viral RNAs between the nucleus and the cytoplasm. HIV-1 Rev interacts with CRM1 in the late phase of HIV-1 replication to promote nuclear export of unspliced and single spliced HIV-1 transcripts. However, the knowledge of cellular factors that are involved in the CRM1-dependent viral RNA nuclear export remains inadequate. Here, we identified that ANP32A and ANP32B mediate the export of unspliced or partially spliced viral mRNA via interacting with Rev and CRM1. We found that double, but not single, knockout of ANP32A and ANP32B, significantly decreased the expression of gag protein. Reconstitution of either ANP32A or ANP32B restored the viral production equally. Disruption of both ANP32A and ANP32B expression led to a dramatic accumulation of unspliced viral mRNA in the nucleus. We further identified that ANP32A and ANP32B interact with both Rev and CRM1 to promote RNA transport and that this function is Rev/RRE-dependent, but not CTE-dependent. Together our data suggests that ANP32A and ANP32B are required for HIV-1 unspliced RNA export in the Rev-CRM1 pathway.Author summaryPosttranscriptional regulation of HIV-1 genome is very important for viral protein expression and viral replication. HIV-1 Rev protein bind to RRE structure of viral RNA and interacts with the mammalian nuclear export factor Chromosomal Maintenance 1 (CRM1) in the late phase of HIV-1 replication to promote nuclear export of unspliced and single spliced HIV-1 transcripts. The REV/RRE-CRM1 pathway has been investigated for years and many host factors have been reported to be involved, but the complicated complex and procedure remain largely unknown. Here the authors report that two host proteins, ANP32A and ANP32B, are novel key factors that support export of unspliced and partial spliced viral RNA from the nucleus to the cytosol. ANP32A/B can interact with both Rev and CRM1, and this interaction is necessary for Rev/RRE-CRM1 dependent viral RNA export. These results suggest that ANP32A and ANP32B are important in viral replication and could be potential targets for novel antiviral strategy.
Yujie Wang; Haili Zhang; Lei Na; Cheng Du; Zhenyu Zhang; Yong-Hui Zheng; Xiaojun Wang. ANP32A and ANP32B are key factors in the Rev dependent CRM1 pathway for nuclear export of HIV-1 unspliced mRNA. 2019, 559096 .
AMA StyleYujie Wang, Haili Zhang, Lei Na, Cheng Du, Zhenyu Zhang, Yong-Hui Zheng, Xiaojun Wang. ANP32A and ANP32B are key factors in the Rev dependent CRM1 pathway for nuclear export of HIV-1 unspliced mRNA. . 2019; ():559096.
Chicago/Turabian StyleYujie Wang; Haili Zhang; Lei Na; Cheng Du; Zhenyu Zhang; Yong-Hui Zheng; Xiaojun Wang. 2019. "ANP32A and ANP32B are key factors in the Rev dependent CRM1 pathway for nuclear export of HIV-1 unspliced mRNA." , no. : 559096.
MLV glycoGag not only enhances MLV replication but also increases HIV-1 infectivity similarly as Nef. Recent studies have discovered that both glycoGag and Nef antagonize a novel host restriction factor Ser5 and promote viral replication. Compared to Nef, the glycoGag antagonism of Ser5 is still poorly understood. MLV glycoGag is a transmembrane version of the structural Gag protein with an extra 88-amino-acid leader region that determines its activity. We now show that glycoGag interacts with Ser5 in live cells and internalizes Ser5 via receptor-mediated endocytosis. Ser5 is polyubiquitinated and relocalized to endosomes and lysosomes for massive destruction. In addition to the previously identified tyrosine-based sorting signal, we find two more important residues for Ser5 relocalization and downregulation. We also find that the Ser5 sensitivity to glycoGag is conserved in the SERINC family. Together, our findings highlight the important role of endosome/lysosome pathway in the enhancement of viral replication by viral proteins.
Sunan Li; Iqbal Ahmad; Jing Shi; Bin Wang; Changqing Yu; Lixin Zhang; Yong-Hui Zheng. Murine Leukemia Virus Glycosylated Gag Reduces Murine SERINC5 Protein Expression at Steady-State Levels via the Endosome/Lysosome Pathway to Counteract SERINC5 Antiretroviral Activity. Journal of Virology 2019, 93, 1 .
AMA StyleSunan Li, Iqbal Ahmad, Jing Shi, Bin Wang, Changqing Yu, Lixin Zhang, Yong-Hui Zheng. Murine Leukemia Virus Glycosylated Gag Reduces Murine SERINC5 Protein Expression at Steady-State Levels via the Endosome/Lysosome Pathway to Counteract SERINC5 Antiretroviral Activity. Journal of Virology. 2019; 93 (2):1.
Chicago/Turabian StyleSunan Li; Iqbal Ahmad; Jing Shi; Bin Wang; Changqing Yu; Lixin Zhang; Yong-Hui Zheng. 2019. "Murine Leukemia Virus Glycosylated Gag Reduces Murine SERINC5 Protein Expression at Steady-State Levels via the Endosome/Lysosome Pathway to Counteract SERINC5 Antiretroviral Activity." Journal of Virology 93, no. 2: 1.
The primate lentiviral accessory protein Nef downregulates CD4 and major histocompatibility complex class I (MHC-I) from the cell surface via independent endosomal trafficking pathways to promote viral pathogenesis. In addition, Nef antagonizes a novel restriction factor, SERINC5 (Ser5), to increase viral infectivity. To explore the molecular mechanism of Ser5 antagonism by Nef, we determined how Nef affects Ser5 expression and intracellular trafficking in comparison to CD4 and MHC-I. We confirm that Nef excludes Ser5 from human immunodeficiency virus type 1 (HIV-1) virions by downregulating its cell surface expression via similar functional motifs required for CD4 downregulation. We find that Nef decreases both Ser5 and CD4 expression at steady-state levels, which are rescued by NH 4 Cl or bafilomycin A1 treatment. Nef binding to Ser5 was detected in living cells using a bimolecular fluorescence complementation assay, where Nef membrane association is required for interaction. In addition, Nef triggers rapid Ser5 internalization via receptor-mediated endocytosis and relocalizes Ser5 to Rab5 + early, Rab7 + late, and Rab11 + recycling endosomes. Manipulation of AP-2, Rab5, Rab7, and Rab11 expression levels affects the Nef-dependent Ser5 and CD4 downregulation. Moreover, although Nef does not promote Ser5 polyubiquitination, Ser5 downregulation relies on the ubiquitination pathway, and both K48- and K63-specific ubiquitin linkages are required for the downregulation. Finally, Nef promotes Ser5 colocalization with LAMP1, which is enhanced by bafilomycin A1 treatment, suggesting that Ser5 is targeted to lysosomes for destruction. We conclude that Nef uses a similar mechanism to downregulate Ser5 and CD4, which sorts Ser5 into a point-of-no-return degradative pathway to counteract its restriction. IMPORTANCE Human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) express an accessory protein called Nef to promote viral pathogenesis. Nef drives immune escape in vivo through downregulation of CD4 and MHC-I from the host cell surface. Recently, Nef was reported to counteract a novel host restriction factor, Ser5, to increase viral infectivity. Nef downregulates cell surface Ser5, thus preventing its incorporation into virus particles, resulting in disruption of its antiviral activity. Here, we report mechanistic studies of Nef-mediated Ser5 downregulation in comparison to CD4 and MHC-I. We demonstrate that Nef binds directly to Ser5 in living cells and that Nef-Ser5 interaction requires Nef association with the plasma membrane. Subsequently, Nef internalizes Ser5 from the plasma membrane via receptor-mediated endocytosis, and targets ubiquitinated Ser5 to endosomes and lysosomes for destruction. Collectively, these results provide new insights into our ongoing understanding of the Nef-Ser5 arms race in HIV-1 infection.
Jing Shi; Ran Xiong; Tao Zhou; Peiyi Su; Xihe Zhang; Xusheng Qiu; Hongmei Li; Sunan Li; Changqing Yu; Bin Wang; Chan Ding; Thomas E. Smithgall; Yong-Hui Zheng. HIV-1 Nef Antagonizes SERINC5 Restriction by Downregulation of SERINC5 via the Endosome/Lysosome System. Journal of Virology 2018, 92, e00196-18 .
AMA StyleJing Shi, Ran Xiong, Tao Zhou, Peiyi Su, Xihe Zhang, Xusheng Qiu, Hongmei Li, Sunan Li, Changqing Yu, Bin Wang, Chan Ding, Thomas E. Smithgall, Yong-Hui Zheng. HIV-1 Nef Antagonizes SERINC5 Restriction by Downregulation of SERINC5 via the Endosome/Lysosome System. Journal of Virology. 2018; 92 (11):e00196-18.
Chicago/Turabian StyleJing Shi; Ran Xiong; Tao Zhou; Peiyi Su; Xihe Zhang; Xusheng Qiu; Hongmei Li; Sunan Li; Changqing Yu; Bin Wang; Chan Ding; Thomas E. Smithgall; Yong-Hui Zheng. 2018. "HIV-1 Nef Antagonizes SERINC5 Restriction by Downregulation of SERINC5 via the Endosome/Lysosome System." Journal of Virology 92, no. 11: e00196-18.
Innate immunity provides an immediate defense against infection after host cells sense danger signals from microbes. Endoplasmic reticulum (ER) stress arises from accumulation of misfolded/unfolded proteins when protein load overwhelms the ER folding capacity, which activates the unfolded protein response (UPR) to restore ER homeostasis. Here, we show that a mechanism for antiviral innate immunity is triggered after the ER stress pathway senses viral glycoproteins. When hemagglutinin (HA) glycoproteins from influenza A virus (IAV) are expressed in cells, ER stress is induced, resulting in rapid HA degradation via proteasomes. The ER-associated protein degradation (ERAD) pathway, an important UPR function for destruction of aberrant proteins, mediates HA degradation. Three class I α-mannosidases were identified to play a critical role in the degradation process, including EDEM1, EDEM2, and ERManI. HA degradation requires either ERManI enzymatic activity or EDEM1/EDEM2 enzymatic activity when ERManI is not expressed, indicating that demannosylation is a critical step for HA degradation. Silencing of EDEM1, EDEM2, and ERManI strongly increases HA expression and promotes IAV replication. Thus, the ER stress pathway senses influenza HA as “nonself” or misfolded protein and sorts HA to ERAD for degradation, resulting in inhibition of IAV replication. IMPORTANCE Viral nucleic acids are recognized as important inducers of innate antiviral immune responses that are sensed by multiple classes of sensors, but other inducers and sensors of viral innate immunity need to be identified and characterized. Here, we used IAV to investigate how host innate immunity is activated. We found that IAV HA glycoproteins induce ER stress, resulting in HA degradation via ERAD and consequent inhibition of IAV replication. In addition, we have identified three class I α-mannosidases, EDEM1, EDEM2, and ERManI, which play a critical role in initiating HA degradation. Knockdown of these proteins substantially increases HA expression and IAV replication. The enzymatic activities and joint actions of these mannosidases are required for this antiviral activity. Our results suggest that viral glycoproteins induce a strong innate antiviral response through activating the ER stress pathway during viral infection.
Dylan Frabutt; Bin Wang; Sana Riaz; Richard C. Schwartz; Yong-Hui Zheng. Innate Sensing of Influenza A Virus Hemagglutinin Glycoproteins by the Host Endoplasmic Reticulum (ER) Stress Pathway Triggers a Potent Antiviral Response via ER-Associated Protein Degradation. Journal of Virology 2018, 92, e01690-17 .
AMA StyleDylan Frabutt, Bin Wang, Sana Riaz, Richard C. Schwartz, Yong-Hui Zheng. Innate Sensing of Influenza A Virus Hemagglutinin Glycoproteins by the Host Endoplasmic Reticulum (ER) Stress Pathway Triggers a Potent Antiviral Response via ER-Associated Protein Degradation. Journal of Virology. 2018; 92 (1):e01690-17.
Chicago/Turabian StyleDylan Frabutt; Bin Wang; Sana Riaz; Richard C. Schwartz; Yong-Hui Zheng. 2018. "Innate Sensing of Influenza A Virus Hemagglutinin Glycoproteins by the Host Endoplasmic Reticulum (ER) Stress Pathway Triggers a Potent Antiviral Response via ER-Associated Protein Degradation." Journal of Virology 92, no. 1: e01690-17.
Among the five serine incorporator (SERINC) family members, SERINC5 (Ser5) was reported to strongly inhibit HIV-1 replication, which is counteracted by Nef. Ser5 produces 5 alternatively spliced isoforms: Ser5-001 has 10 putative transmembrane domains, whereas Ser5-004, -005, -008a, and -008b do not have the last one. Here, we confirmed the strong Ser5 anti-HIV-1 activity and investigated its isoforms' expression and antiviral activities. It was found that Ser5-001 transcripts were detected at least 10-fold more than the other isoforms by real-time quantitative PCR. When Ser5-001 and its two isoforms Ser5-005 and Ser5-008a were expressed from the same mammalian expression vector, only Ser5-001 was stably expressed, whereas the others were poorly expressed due to rapid degradation. In addition, unlike the other isoforms, which are located mainly in the cytoplasm, Ser5-001 is localized primarily to the plasma membrane. To map the critical determinant, Ser5 mutants bearing C-terminal deletions were created. It was found that the 10th transmembrane domain is required for Ser5 stable expression and plasma membrane localization. As expected, only Ser5-001 strongly inhibits HIV-1 infectivity, whereas the other Ser5 isoforms and mutants that do not have the 10th transmembrane domain show very poor activity. It was also observed that the Nef counteractive activity could be easily saturated by Ser5 overexpression. Thus, we conclude that Ser5-001 is the predominant antiviral isoform that restricts HIV-1, and the 10th transmembrane domain plays a critical role in this process by regulating its protein stability and plasma membrane targeting. IMPORTANCE Human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) express a small protein, Nef, to enhance viral pathogenesis in vivo . Nef has an important in vitro function, which is to make virus particles more infectious, but the mechanism has been unclear. Recently, Nef was reported to counteract a novel anti-HIV host protein, SERINC5 (Ser5). Ser5 has five alternatively spliced isoforms, Ser5-001, -004, -005, -008a, and -008b, and only Ser5-001 has an extra C-terminal transmembrane domain. We now show that the Ser5-001 transcripts are produced at least 10-fold more than the others, and only Ser5-001 produces stable proteins that are targeted to the plasma membrane. Importantly, only Ser5-001 shows strong anti-HIV-1 activity. We further demonstrate that the extra transmembrane domain is required for Ser5 stable expression and plasma membrane localization. These results suggest that plasma membrane localization is required for Ser5 antiviral activity, and Ser5-001 is the predominant isoform that contributes to the activity.
Xianfeng Zhang; Tao Zhou; Jie Yang; Yumei Lin; Jing Shi; Xihe Zhang; Dylan Frabutt; Xiangwei Zeng; Sunan Li; Patrick J. Venta; Yong-Hui Zheng. Identification of SERINC5-001 as the Predominant Spliced Isoform for HIV-1 Restriction. Journal of Virology 2017, 91, e00137-17 .
AMA StyleXianfeng Zhang, Tao Zhou, Jie Yang, Yumei Lin, Jing Shi, Xihe Zhang, Dylan Frabutt, Xiangwei Zeng, Sunan Li, Patrick J. Venta, Yong-Hui Zheng. Identification of SERINC5-001 as the Predominant Spliced Isoform for HIV-1 Restriction. Journal of Virology. 2017; 91 (10):e00137-17.
Chicago/Turabian StyleXianfeng Zhang; Tao Zhou; Jie Yang; Yumei Lin; Jing Shi; Xihe Zhang; Dylan Frabutt; Xiangwei Zeng; Sunan Li; Patrick J. Venta; Yong-Hui Zheng. 2017. "Identification of SERINC5-001 as the Predominant Spliced Isoform for HIV-1 Restriction." Journal of Virology 91, no. 10: e00137-17.
The Ebola virus (EBOV) trimeric envelope glycoprotein (GP) precursors are cleaved into the receptor-binding GP1 and the fusion-mediating GP2 subunits and incorporated into virions to initiate infection. GP1 and GP2 form heterodimers that have 15 or two N-glycosylation sites (NGSs), respectively. Here we investigated the mechanism of how N-glycosylation contributes to GP expression, maturation, and function. As reported before, we found that, although GP1 NGSs are not critical, the two GP2 NGSs, Asn563 and Asn618, are essential for GP function. Further analysis uncovered that Asn563 and Asn618 regulate GP processing, demannosylation, oligomerization, and conformation. Consequently, these two NGSs are required for GP incorporation into EBOV-like particles and HIV type 1 (HIV-1) pseudovirions and determine viral transduction efficiency. Using CRISPR/Cas9 technology, we knocked out the two classical endoplasmic reticulum chaperones calnexin (CNX) and/or calreticulin (CRT) and found that both CNX and CRT increase GP expression. Nevertheless, NGSs are not required for the GP interaction with CNX or CRT. Together, we conclude that, although Asn563 and Asn618 are not required for EBOV GP expression, they synergistically regulate its maturation, which determines its functionality.
Bin Wang; Yujie Wang; Dylan A. Frabutt; Xihe Zhang; Xiaoyu Yao; Dan Hu; Zhuo Zhang; Chaonan Liu; Shimin Zheng; Shi-Hua Xiang; Yong-Hui Zheng. Mechanistic understanding of N-glycosylation in Ebola virus glycoprotein maturation and function. Journal of Biological Chemistry 2017, 292, 5860 -5870.
AMA StyleBin Wang, Yujie Wang, Dylan A. Frabutt, Xihe Zhang, Xiaoyu Yao, Dan Hu, Zhuo Zhang, Chaonan Liu, Shimin Zheng, Shi-Hua Xiang, Yong-Hui Zheng. Mechanistic understanding of N-glycosylation in Ebola virus glycoprotein maturation and function. Journal of Biological Chemistry. 2017; 292 (14):5860-5870.
Chicago/Turabian StyleBin Wang; Yujie Wang; Dylan A. Frabutt; Xihe Zhang; Xiaoyu Yao; Dan Hu; Zhuo Zhang; Chaonan Liu; Shimin Zheng; Shi-Hua Xiang; Yong-Hui Zheng. 2017. "Mechanistic understanding of N-glycosylation in Ebola virus glycoprotein maturation and function." Journal of Biological Chemistry 292, no. 14: 5860-5870.
Enveloped viruses represent a significant category of pathogens that cause serious diseases in animals. These viruses express envelope glycoproteins that are singularly important during the infection of host cells by mediating fusion between the viral envelope and host cell membranes. Despite low homology at protein levels, three classes of viral fusion proteins have, as of yet, been identified based on structural similarities. Their incorporation into viral particles is dependent upon their proper sub-cellular localization after being expressed and folded properly in the endoplasmic reticulum (ER). However, viral protein expression can cause stress in the ER, and host cells respond to alleviate the ER stress in the form of the unfolded protein response (UPR); the effects of which have been observed to potentiate or inhibit viral infection. One important arm of UPR is to elevate the capacity of the ER-associated protein degradation (ERAD) pathway, which is comprised of host quality control machinery that ensures proper protein folding. In this review, we provide relevant details regarding viral envelope glycoproteins, UPR, ERAD, and their interactions in host cells.
Dylan A. Frabutt; Yong-Hui Zheng. Arms Race between Enveloped Viruses and the Host ERAD Machinery. Viruses 2016, 8, 255 .
AMA StyleDylan A. Frabutt, Yong-Hui Zheng. Arms Race between Enveloped Viruses and the Host ERAD Machinery. Viruses. 2016; 8 (9):255.
Chicago/Turabian StyleDylan A. Frabutt; Yong-Hui Zheng. 2016. "Arms Race between Enveloped Viruses and the Host ERAD Machinery." Viruses 8, no. 9: 255.
Vpr enhances HIV-1 replication in macrophages and dendritic cells, as well as the human CD4+ CEM.NKR T cell line. Recently, Vpr was reported to increase HIV-1 Env expression in macrophages. Here, we report that Vpr also increases HIV-1 Env expression in dendritic cells and CEM.NKR cells. The Vpr activity depends on its N-terminal region, which was disrupted by a single A30L mutation. Env was rapidly degraded in the absence of Vpr, which was blocked by the ERAD pathway inhibitor kifunesine or the lysosome inhibitor Bafilomycin. As2O3 or PK11195, which reportedly enhances HIV-1 Env folding, also blocked the Env degradation in CEM.NKR cells. Thus, these results not only identify Env as a primary target for Vpr to boost HIV-1 replication, but also suggest that Vpr likely promotes Env folding in the ER, which is otherwise misfolded and targeted by the ERAD pathway to lysosomes for degradation.
Xianfeng Zhang; Tao Zhou; Dylan Frabutt; Yong-Hui Zheng. HIV-1 Vpr increases Env expression by preventing Env from endoplasmic reticulum-associated protein degradation (ERAD). Virology 2016, 496, 194 -202.
AMA StyleXianfeng Zhang, Tao Zhou, Dylan Frabutt, Yong-Hui Zheng. HIV-1 Vpr increases Env expression by preventing Env from endoplasmic reticulum-associated protein degradation (ERAD). Virology. 2016; 496 ():194-202.
Chicago/Turabian StyleXianfeng Zhang; Tao Zhou; Dylan Frabutt; Yong-Hui Zheng. 2016. "HIV-1 Vpr increases Env expression by preventing Env from endoplasmic reticulum-associated protein degradation (ERAD)." Virology 496, no. : 194-202.