title: Retroviruses pseudotyped with the severe acute respiratory syndrome coronavirus spike protein efficiently infect cells expressing angiotensin-converting enzyme 2.
abstract: Infection of receptor-bearing cells by coronaviruses is mediated by their spike (S) proteins. The coronavirus (SARS-CoV) that causes severe acute respiratory syndrome (SARS) infects cells expressing the receptor angiotensin-converting enzyme 2 (ACE2). Here we show that codon optimization of the SARS-CoV S-protein gene substantially enhanced S-protein expression. We also found that two retroviruses, simian immunodeficiency virus (SIV) and murine leukemia virus, both expressing green fluorescent protein and pseudotyped with SARS-CoV S protein or S-protein variants, efficiently infected HEK293T cells stably expressing ACE2. Infection mediated by an S-protein variant whose cytoplasmic domain had been truncated and altered to include a fragment of the cytoplasmic tail of the human immunodeficiency virus type 1 envelope glycoprotein was, in both cases, substantially more efficient than that mediated by wild-type S protein. Using S-protein-pseudotyped SIV, we found that the enzymatic activity of ACE2 made no contribution to S-protein-mediated infection. Finally, we show that a soluble and catalytically inactive form of ACE2 potently blocked infection by S-protein-pseudotyped retrovirus and by SARS-CoV. These results permit studies of SARS-CoV entry inhibitors without the use of live virus and suggest a candidate therapy for SARS.
authors.affiliation: Partners AIDS Research Center, Brigham and Women's Hospital, and Department of Medicine, Harvard Medical School, Boston, MA, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Moore
authors.forename: Tatyana
authors.initials: T
authors.lastname: Dorfman
authors.forename: Wenhui
authors.initials: W
authors.lastname: Li
authors.forename: Swee Kee
authors.initials: SK
authors.lastname: Wong
authors.forename: Yanhan
authors.initials: Y
authors.lastname: Li
authors.forename: Jens H
authors.initials: JH
authors.lastname: Kuhn
authors.forename: James
authors.initials: J
authors.lastname: Coderre
authors.forename: Natalya
authors.initials: N
authors.lastname: Vasilieva
authors.forename: Zhongchao
authors.initials: Z
authors.lastname: Han
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Greenough
authors.forename: Michael
authors.initials: M
authors.lastname: Farzan
authors.forename: Hyeryun
authors.initials: H
authors.lastname: Choe
mesh_terms: D000595:Amino Acid Sequence; D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D002268:Carboxypeptidases / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D007703:Peptidyl-Dipeptidase A; D011991:Receptors, Virus / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism*; D014771:Virion / Q000737:chemistry / Q000378:metabolism; D014779:Virus Replication
chemical_list: D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus; D002268:Carboxypeptidases; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; D000085962:Angiotensin-Converting Enzyme 2