Effect of polymorphism in Rhinolophus affinis ACE2 on entry of SARS-CoV-2 related bat coronaviruses.
Effect of polymorphism in Rhinolophus affinis ACE2 on entry of SARS-CoV-2 related bat coronaviruses.
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DOI：
10.1371/journal.ppat.1011116
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发表时间：
2023-01
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
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蝙蝠冠状病毒RaTG 13与SARS-CoV-2具有约96.2%的核苷酸序列同一性，并以人和菊头蝠（Rhinolophus affinis，Ra）血管紧张素转换酶2（ACE 2）作为进入受体。是否存在除R.对RaTG 13感染敏感的近亲仍然难以捉摸。在这里，我们表明，在18个不同的蝙蝠ACE 2测试，只有RaACE 2是高度敏感的转导RaTG 13 S假病毒，表明蝙蝠物种窝藏RaTG 13可能是非常有限的。RaACE 2有7个多态性变体，RA-01至RA-07，它们对RaTG 13 S假病毒体转导显示出不同的亲和性。序列分析和突变分析表明，RaACE 2的34，38和83位残基可能在与RaTG 13 S蛋白的相互作用中发挥关键作用。值得注意的是，RaACE 2多态性在结合、膜融合和假病毒进入方面对SARS-CoV-2和几种SARS-CoV-2相关CoV（SC2 r-CoV）（包括BANAL-20-52和BANAL-20-236）的S蛋白具有最小的影响。进一步的诱变分析鉴定了S蛋白中对不同RaACE 2变体和穿山甲ACE 2（pACE 2）识别至关重要的残基501和505，表明RaTG 13可能没有很好地适应R.近亲蝙蝠虽然RaTG 13 S蛋白中的单个D501 N和H505 Y变化显著增强了感染性并使不同RaACE 2变体之间的易感性差异最小化，但SARS-CoV-2 S蛋白中的N501 D取代显示了RaACE 2变体之间转导效率的显著差异，其中几种RaACE 2变体的感染性显著降低。最后，RaTG 13 S蛋白中的T372 A取代不仅显著增加了对所有RaACE 2变体的感染性，而且显著增强了对包括R. sinicus YN、中华弯角叶蝉R. pearsonii和R. ferrumeiqunum。然而，T372 A突变体对来自用BANAL-20-52 S免疫的小鼠的中和血清的敏感性高约4倍，这表明T372优于A372的免疫逃避能力可能有助于T372在蝙蝠CoV中优于A372的天然选择性优势。总之，我们的研究有助于更好地了解冠状病毒的进入，疫苗设计和进化。蝙蝠冠状病毒RaTG 13具有人畜共患传播的潜力，S蛋白与ACE 2的相互作用在其中起着重要作用。RaTG 13使用RaACE 2作为进入受体，并且存在至少七种多态性RaACE 2变体。RaACE 2多态性对RaTG 13和其他SARS-CoV-2相关蝙蝠CoV（SC2 r-CoV）进入的影响尚未确定。在本研究中，我们发现RaTG 13 S假病毒体仅在中国西南和东南亚常见的18种蝙蝠中高效地表达RaACE 2，这表明可能存在有限的蝙蝠物种，包括R.对感染RaTG 13病毒敏感的近亲。RaACE 2多态性对RaTG 13的进入具有显著影响，而它对SARS-CoV-2和SC2 r-CoV如BANAL-20-52、BANAL-20-236和pangolin-CoV GD的进入显示出最小的影响。进一步的序列和诱变分析鉴定了RaACE 2中的残基34、38和83以及S蛋白中对S蛋白和受体相互作用至关重要的残基501和505，表明RaTG 13可能没有很好地适应R。近亲蝙蝠我们还发现，T372 A在RaTG 13 S取代显着增强进入所有RaACE 2变体和几个蝙蝠ACE 2，和免疫逃避可能有助于在蝙蝠S蛋白的T372超过A372的自然选择。总之，我们的工作提供了RaACE 2多态性对RaTG 13和其他SC2 r-CoV进入的影响的详细分析，并更好地理解CoV的进入和进化。
Bat coronavirus RaTG13 shares about 96.2% nucleotide sequence identity with that of SARS-CoV-2 and uses human and Rhinolophus affinis (Ra) angiotensin-converting enzyme 2 (ACE2) as entry receptors. Whether there are bat species other than R. affinis susceptible to RaTG13 infection remains elusive. Here, we show that, among 18 different bat ACE2s tested, only RaACE2 is highly susceptible to transduction by RaTG13 S pseudovirions, indicating that the bat species harboring RaTG13 might be very limited. RaACE2 has seven polymorphic variants, RA-01 to RA-07, and they show different susceptibilities to RaTG13 S pseudovirions transduction. Sequence and mutagenesis analyses reveal that residues 34, 38, and 83 in RaACE2 might play critical roles in interaction with the RaTG13 S protein. Of note, RaACE2 polymorphisms have minimal effect on S proteins of SARS-CoV-2 and several SARS-CoV-2 related CoVs (SC2r-CoVs) including BANAL-20-52 and BANAL-20-236 in terms of binding, membrane fusion, and pseudovirus entry. Further mutagenesis analyses identify residues 501 and 505 in S proteins critical for the recognition of different RaACE2 variants and pangolin ACE2 (pACE2), indicating that RaTG13 might have not been well adapted to R. affinis bats. While single D501N and H505Y changes in RaTG13 S protein significantly enhance the infectivity and minimize the difference in susceptibility among different RaACE2 variants, an N501D substitution in SARS-CoV-2 S protein displays marked disparity in transduction efficiencies among RaACE2 variants with a significant reduction in infectivity on several RaACE2 variants. Finally, a T372A substitution in RaTG13 S protein not only significantly increases infectivity on all RaACE2 variants, but also markedly enhances entry on several bat ACE2s including R. sinicus YN, R. pearsonii, and R. ferrumeiqunum. However, the T372A mutant is about 4-fold more sensitive to neutralizing sera from mice immunized with BANAL-20-52 S, suggesting that the better immune evasion ability of T372 over A372 might contribute to the natural selective advantage of T372 over A372 among bat CoVs. Together, our study aids a better understanding of coronavirus entry, vaccine design, and evolution. Bat CoV RaTG13 has zoonotic transmission potential, and interactions between S protein and ACE2 play essential roles. RaTG13 uses RaACE2 as an entry receptor, and there are at least seven polymorphic RaACE2 variants. The effect of RaACE2 polymorphism on entry of RaTG13 and other SARS-CoV-2 related bat CoVs (SC2r-CoVs) has not been determined. In this study, we find that RaTG13 S pseudovirions only highly efficiently transduce cells expressing RaACE2 among available 18 bat species commonly habituating in Southwestern China and Southeastern Asia, suggesting that there might be limited bat species including R. affinis susceptible to infection of RaTG13 virus. The RaACE2 polymorphism has a marked effect on the entry of RaTG13, whereas it shows a minimal effect on the entry of SARS-CoV-2 and SC2r-CoVs like BANAL-20-52, BANAL-20-236, and pangolin-CoV GD. Further sequence and mutagenesis analyses identify residues 34, 38, and 83 in RaACE2 and residues 501 and 505 in the S proteins critical for S protein and receptor interactions, indicating that RaTG13 might have not been well adapted to R. affinis bats. We also find that T372A substitution in RaTG13 S significantly enhances entry on all RaACE2 variants and several bat ACE2s, and immune evasion might contribute to the natural selection of T372 over A372 in bat S proteins. Altogether, our work provides detailed analyses of the effect of RaACE2 polymorphism on the entry of RaTG13 and other SC2r-CoVs and a better understanding of the entry and evolution of CoVs.
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DOI：
10.1126/science.abc4730
发表时间：
2020-09-25
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gu H;Chen Q;Yang G;He L;Fan H;Deng YQ;Wang Y;Teng Y;Zhao Z;Cui Y;Li Y;Li XF;Li J;Zhang NN;Yang X;Chen S;Guo Y;Zhao G;Wang X;Luo DY;Wang H;Yang X;Li Y;Han G;He Y;Zhou X;Geng S;Sheng X;Jiang S;Sun S;Qin CF;Zhou Y
通讯作者：
Zhou Y
DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
影响因子：
5.4
作者：
Ren W;Zhu Y;Lan J;Chen H;Wang Y;Shi H;Feng F;Chen DY;Close B;Zhao X;Wu J;Tian B;Yuan Z;Zhou D;Saeed M;Wang X;Zhang R;Ding Q
通讯作者：
Ding Q
影响因子：
28.3
作者：
Gorbalenya, Alexander E.;Baker, Susan C.;Ziebuhr, John
通讯作者：
Ziebuhr, John
影响因子：
3.7
作者：
Hattori, Takanari;Saito, Takeshi;Okuya, Kosuke;Takahashi, Yuji;Miyamoto, Hiroko;Kajihara, Masahiro;Igarashi, Manabu;Takadaa, Ayato
通讯作者：
Takadaa, Ayato