title: Further Evidence for Bats as the Evolutionary Source of Middle East Respiratory Syndrome Coronavirus.
abstract: The evolutionary origins of Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) are unknown. Current evidence suggests that insectivorous bats are likely to be the original source, as several 2c CoVs have been described from various species in the familyVespertilionidaeHere, we describe a MERS-like CoV identified from aPipistrellus cf. hesperidusbat sampled in Uganda (strain PREDICT/PDF-2180), further supporting the hypothesis that bats are the evolutionary source of MERS-CoV. Phylogenetic analysis showed that PREDICT/PDF-2180 is closely related to MERS-CoV across much of its genome, consistent with a common ancestry; however, the spike protein was highly divergent (46% amino acid identity), suggesting that the two viruses may have different receptor binding properties. Indeed, several amino acid substitutions were identified in key binding residues that were predicted to block PREDICT/PDF-2180 from attaching to the MERS-CoV DPP4 receptor. To experimentally test this hypothesis, an infectious MERS-CoV clone expressing the PREDICT/PDF-2180 spike protein was generated. Recombinant viruses derived from the clone were replication competent but unable to spread and establish new infections in Vero cells or primary human airway epithelial cells. Our findings suggest that PREDICT/PDF-2180 is unlikely to pose a zoonotic threat. Recombination in the S1 subunit of the spike gene was identified as the primary mechanism driving variation in the spike phenotype and was likely one of the critical steps in the evolution and emergence of MERS-CoV in humans.IMPORTANCEGlobal surveillance efforts for undiscovered viruses are an important component of pandemic prevention initiatives. These surveys can be useful for finding novel viruses and for gaining insights into the ecological and evolutionary factors driving viral diversity; however, finding a viral sequence is not sufficient to determine whether it can infect people (i.e., poses a zoonotic threat). Here, we investigated the specific zoonotic risk of a MERS-like coronavirus (PREDICT/PDF-2180) identified in a bat from Uganda and showed that, despite being closely related to MERS-CoV, it is unlikely to pose a threat to humans. We suggest that this approach constitutes an appropriate strategy for beginning to determine the zoonotic potential of wildlife viruses. By showing that PREDICT/PDF-2180 does not infect cells that express the functional receptor for MERS-CoV, we further show that recombination was likely to be the critical step that allowed MERS to emerge in humans.
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA sja2127@cumc.columbia.edu.|Department of Epidemiology, Mailman School of Public Health, Columbia University, New York, New York, USA.|EcoHealth Alliance, New York, New York, USA.
authors.forename: S J
authors.initials: SJ
authors.lastname: Anthony
authors.affiliation: One Health Institute and Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, California, USA.
authors.forename: K
authors.initials: K
authors.lastname: Gilardi
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: V D
authors.initials: VD
authors.lastname: Menachery
authors.affiliation: One Health Institute and Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, California, USA.
authors.forename: T
authors.initials: T
authors.lastname: Goldstein
authors.affiliation: Gorilla Doctors, c/o MGVP, Inc., Davis, California, USA.
authors.forename: B
authors.initials: B
authors.lastname: Ssebide
authors.affiliation: Gorilla Doctors, c/o MGVP, Inc., Davis, California, USA.
authors.forename: R
authors.initials: R
authors.lastname: Mbabazi
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: I
authors.initials: I
authors.lastname: Navarrete-Macias
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.|EcoHealth Alliance, New York, New York, USA.
authors.forename: E
authors.initials: E
authors.lastname: Liang
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: H
authors.initials: H
authors.lastname: Wells
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: A
authors.initials: A
authors.lastname: Hicks
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: A
authors.initials: A
authors.lastname: Petrosov
authors.affiliation: Makerere University Walter Reed Project, Kampala, Uganda.|Makerere University, College of Veterinary Medicine, Kampala, Uganda.
authors.forename: D K
authors.initials: DK
authors.lastname: Byarugaba
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: K
authors.initials: K
authors.lastname: Debbink
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: K H
authors.initials: KH
authors.lastname: Dinnon
authors.affiliation: Department of Epidemiology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: T
authors.initials: T
authors.lastname: Scobey
authors.affiliation: Department of Cell Biology and Physiology and Marsico Lung Institute/Cystic Fibrosis Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: S H
authors.initials: SH
authors.lastname: Randell
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: B L
authors.initials: BL
authors.lastname: Yount
authors.affiliation: One Health Institute and Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, California, USA.|Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: M
authors.initials: M
authors.lastname: Cranfield
authors.affiliation: One Health Institute and Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, California, USA.
authors.forename: C K
authors.initials: CK
authors.lastname: Johnson
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: R S
authors.initials: RS
authors.lastname: Baric
authors.affiliation: Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA.|Department of Epidemiology, Mailman School of Public Health, Columbia University, New York, New York, USA.
authors.forename: W I
authors.initials: WI
authors.lastname: Lipkin
authors.affiliation: One Health Institute and Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, California, USA.
authors.forename: J A K
authors.initials: JA
authors.lastname: Mazet
mesh_terms: D000818:Animals; D002685:Chiroptera / Q000821:virology*; D019143:Evolution, Molecular; D016679:Genome, Viral; D065207:Middle East Respiratory Syndrome Coronavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D010802:Phylogeny; D011991:Receptors, Virus / Q000378:metabolism; D064370:Spike Glycoprotein, Coronavirus / Q000235:genetics / Q000378:metabolism; D026801:Synteny; D014454:Uganda; D053585:Virus Attachment
chemical_list: D011991:Receptors, Virus; D064370:Spike Glycoprotein, Coronavirus
keywords: MERS coronavirus; Uganda; bat; spike; zoonoses