Expression cloning of functional receptor used by SARS coronavirus.
We have expressed a series of truncated spike (S) glycoproteins of SARS-CoV and found that the N-terminus 14-502 residuals were sufficient to bind to SARS-CoV susceptible Vero E6 cells. With this soluble S protein fragment as an affinity ligand, we screened HeLa cells transduced with retroviral cDNA library from Vero E6 cells and obtained a HeLa cell clone which could bind with the S protein. This cell clone was susceptible to HIV/SARS pseudovirus infection and the presence of a functional receptor for S protein in this cell clone was confirmed by the cell-cell fusion assay. Further studies showed the susceptibility of this cell was due to the expression of endogenous angiotensin-converting enzyme 2 (ACE2) which was activated by inserted LTR from retroviral vector used for expression cloning. When human ACE2 cDNA was transduced into NIH3T3 cells, the ACE2 expressing NIH3T3 cells could be infected with HIV/SARS pseudovirus. These data clearly demonstrated that ACE2 was the functional receptor for SARS-CoV.
Wang
Peigang
P
Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, PR China.
Chen
Jian
J
Zheng
Aihua
A
Nie
Yuchun
Y
Shi
Xuanling
X
Wang
Wei
W
Wang
Guangwen
G
Luo
Min
M
Liu
Huijun
H
Tan
Lei
L
Song
Xijun
X
Wang
Zai
Z
Yin
Xiaolei
X
Qu
Xiuxia
X
Wang
Xiaojing
X
Qing
Tingting
T
Ding
Mingxiao
M
Deng
Hongkui
H
D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D002268:Carboxypeptidases / Q000378:metabolism; D002352:Carrier Proteins; D002469:Cell Separation; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D005434:Flow Cytometry; D015723:Gene Library; D006023:Glycoproteins / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D041681:NIH 3T3 Cells; D007703:Peptidyl-Dipeptidase A; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011991:Receptors, Virus / Q000737:chemistry*; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000378:metabolism*; D020079:Terminal Repeat Sequences; D014709:Vero Cells; D014759:Viral Envelope Proteins
D002352:Carrier Proteins; D018076:DNA, Complementary; D006023:Glycoproteins; D008024:Ligands; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D014759:Viral Envelope Proteins; D002268:Carboxypeptidases; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; C000705328:Ace2 protein, mouse; D000085962:Angiotensin-Converting Enzyme 2