Identification of Two Critical Amino Acid Residues of the Severe Acute Respiratory Syndrome Coronavirus Spike Protein for Its Variation in Zoonotic Tropism Transition via a Double Substitution Strategy

Qu, Xiuxia; Hao, Pei; Song, Xiaoyu; Jiang, Shantong; Liu, Yanxia; Wang, Peigang; Rao, Xi; Song, Huai‐Dong; Wang, Shengyue; Zuo, Yu; Zheng, Aihua; Luo, Min; Wang, Hualin; Deng, Fei; Wang, Hanzhong; Hú, Zhìhóng; Ding, Mei; Zhao, Guoping; Deng, Hongkui

doi:10.1074/jbc.m500662200

Cited by 161 publications

(171 citation statements)

References 34 publications

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“…However, RBD Thr487 in the human S protein is replaced by Ser487 in the palm civet and raccoon dog S proteins, and there is no c-methyl group in Ser487. It has also been reported that Ser487 on RBD dramatically reduces the affinity for huACE2 by over 20-fold (W. Li et al, 2005a;Qu et al, 2005). This may explain why pseudoviruses derived from SARS-CoV-like viruses of palm civets and raccoon dogs had lower entry efficiencies into HeLa-rdACE2 cells than human SARS-CoV.…”

Section: Discussionmentioning

confidence: 81%

“…Although their infectivities are very similar, it is possible that, when these four SNPs exist together, the influence on S-ACE2 interaction is limited. However, when another two mutations at residues 344 and 487 are added to the above four SNPs, virus entry efficiencies are inhibited dramatically, despite the hypothesis of Qu et al (2005) and Li (2008) that the inhibition was mainly caused by mutations of residues 479 and 487. However, we need to mention that, apart from the RBD region, other parts of the S genes from the raccoon dog viruses also contain mutations that may have impacts on entry efficiency.…”

Section: Discussionmentioning

confidence: 99%

“…The HeLa-huACE2 and HeLa-pcACE2 cell lines, which stably express ACE2 from human and palm civet, respectively, were constructed previously (Qu et al, 2005). Generation of a cell line stably expressing rdACE2 was established by using a Moloney murine leukemia virus-based pseudotyped system termed pVPack (Stratagene).…”

Section: Methodsmentioning

confidence: 99%

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Angiotensin-converting enzyme 2 (ACE2) from raccoon dog can serve as an efficient receptor for the spike protein of severe acute respiratory syndrome coronavirus

Zhang

Liu

et al. 2009

Journal of General Virology

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Raccoon dog is one of the suspected intermediate hosts of severe acute respiratory syndrome coronavirus (SARS-CoV). In this study, the angiotensin-converting enzyme 2 (ACE2) gene of raccoon dog (rdACE2) was cloned and sequenced. The amino acid sequence of rdACE2 has identities of 99.3, 89.2, 83.9 and 80.4 % to ACE2 proteins from dog, masked palm civet (pcACE2), human (huACE2) and bat, respectively. There are six amino acid changes in rdACE2 compared with huACE2, and four changes compared with pcACE2, within the 18 residues of ACE2 known to make direct contact with the SARS-CoV S protein. A HeLa cell line stably expressing rdACE2 was established; Western blot analyses and an enzyme-activity assay indicated that the cell line expressed ACE2 at a similar level to two previously established cell lines that express ACE2 from human and masked palm civet, respectively. Human immunodeficiency virus-backboned pseudoviruses expressing spike proteins derived from human SARS-CoV or SARS-CoV-like viruses of masked palm civets and raccoon dogs were tested for their entry efficiency into these cell lines. The results showed that rdACE2 is a more efficient receptor for human SARS-CoV, but not for SARS-CoV-like viruses of masked palm civets and raccoon dogs, than huACE2 or pcACE2. This study provides useful data to elucidate the role of raccoon dog in SARS outbreaks.

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Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

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Angiotensin-converting enzyme 2 (ACE2) from raccoon dog can serve as an efficient receptor for the spike protein of severe acute respiratory syndrome coronavirus

Zhang

Liu

et al. 2009

Journal of General Virology

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“…For standardization of the amount of mRNA, expression of GAPDH in each sample was also quantified. All PCRs were performed using an ABI Prism 7000 sequence detector (Perkin-Elmer Applied Biosystems) as described previously (27). Briefly, equivalent amounts of cDNA sample, derived from 50 ng of DNase-digested total RNA, were used for each PCR.…”

Section: Methodsmentioning

confidence: 99%

Claudin-6 and Claudin-9 Function as Additional Coreceptors for Hepatitis C Virus

Zheng

Yuan

et al. 2007

J Virol

186

188

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Hepatitis C virus (HCV) is a global challenge to public health. Several factors have been proven to be critical for HCV entry, including the newly identified claudin-1 (CLDN1). However, the mechanism of HCV entry is still obscure. Presently, among the 20 members of the claudin family identified in humans so far, CLDN1 has been the only member shown to be necessary for HCV entry. Recently, we discovered that Bel7402, an HCV-permissive cell line, does not express CLDN1 but expresses other members of claudin family. Among these claudins, CLDN9 was able to mediate HCV entry just as efficiently as CLDN1. We then examined if other members of the claudin family could mediate entry. We show that CLDN6 and CLDN9, but not CLDN2, CLDN3, CLDN4, CLDN7, CLDN11, CLDN12, CLDN15, CLDN17, and CLDN23, were able to mediate the entry of HCV into target cells. We found that CLDN6 and CLDN9 are expressed in the liver, the primary site of HCV replication. We also showed that CLDN6 and CLDN9, but not CLDN1, are expressed in peripheral blood mononuclear cells, an additional site of HCV replication. Through sequence comparison and mutagenesis studies, we show that residues N38 and V45 in the first extracellular loop (EL1) of CLDN9 are necessary for HCV entry.Hepatitis C virus (HCV) is the major cause of liver cirrhosis and hepatocellular carcinoma worldwide. Approximately 3% of the global population is infected with HCV, and at least 70% develop chronic hepatitis (13,17,32). In patients with chronic HCV infection, about 20% develop liver cirrhosis, about 5% of which go on to develop hepatocellular carcinoma (17). While HCV is generally confined to the liver, there is growing evidence suggesting that HCV can replicate in extrahepatic tissues including peripheral blood mononuclear cells (PBMCs) (4,15,23,24

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“…However, the RBDs among the human 2002-2003 and 2003-2004 and civet SARS-CoV isolates differ in several amino acids (Fig. 1B), some of which determine the receptor adaptation of SARS-CoV from palm civet to human (43,44). It is important to know whether these sequence variations can determine the antigenic epitopes in the RBD and whether the recombinant RBDs can induce Abs with broad cross-neutralizing activity.…”

Section: S Evere Acute Respiratory Syndrome (Sars)mentioning

confidence: 99%

Cross-Neutralization of Human and Palm Civet Severe Acute Respiratory Syndrome Coronaviruses by Antibodies Targeting the Receptor-Binding Domain of Spike Protein

et al. 2006

The Journal of Immunology

112

119

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The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is considered as a protective Ag for vaccine design. We previously demonstrated that the receptor-binding domain (RBD) of S protein contains multiple conformational epitopes (Conf I-VI) that confer the major target of neutralizing Abs. Here we show that the recombinant RBDs derived from the S protein sequences of Tor2, GD03, and SZ3, the representative strains of human 2002–2003 and 2003–2004 SARS-CoV and palm civet SARS-CoV, respectively, induce in the immunized mice and rabbits high titers of cross-neutralizing Abs against pseudoviruses expressing S proteins of Tor2, GD03, and SZ3. We also demonstrate that the Tor2-RBD induced-Conf I-VI mAbs can potently neutralize both human SARS-CoV strains, Tor2 and GD03. However, only the Conf IV-VI, but not Conf I-III mAbs, neutralize civet SARS-CoV strain SZ3. All these mAbs reacted significantly with each of the three RBD variants (Tor2-RBD, GD03-RBD, and SZ3-RBD) that differ at several amino acids. Regardless, the Conf I-IV and VI epitopes were completely disrupted by single-point mutation of the conserved residues in the RBD (e.g., D429A, R441A, or D454A) and the Conf III epitope was significantly affected by E452A or D463A substitution. Interestingly, the Conf V epitope, which may overlap the receptor-binding motif and induce most potent neutralizing Abs, was conserved in these mutants. These data suggest that the major neutralizing epitopes of SARS-CoV have been apparently maintained during cross-species transmission, and that RBD-based vaccines may induce broad protection against both human and animal SARS-CoV variants.

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Identification of Two Critical Amino Acid Residues of the Severe Acute Respiratory Syndrome Coronavirus Spike Protein for Its Variation in Zoonotic Tropism Transition via a Double Substitution Strategy

Cited by 161 publications

References 34 publications

Angiotensin-converting enzyme 2 (ACE2) from raccoon dog can serve as an efficient receptor for the spike protein of severe acute respiratory syndrome coronavirus

Angiotensin-converting enzyme 2 (ACE2) from raccoon dog can serve as an efficient receptor for the spike protein of severe acute respiratory syndrome coronavirus

Claudin-6 and Claudin-9 Function as Additional Coreceptors for Hepatitis C Virus

Cross-Neutralization of Human and Palm Civet Severe Acute Respiratory Syndrome Coronaviruses by Antibodies Targeting the Receptor-Binding Domain of Spike Protein

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