title: Host species restriction of Middle East respiratory syndrome coronavirus through its receptor, dipeptidyl peptidase 4.
abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) emerged in 2012. Recently, the MERS-CoV receptor dipeptidyl peptidase 4 (DPP4) was identified and the specific interaction of the receptor-binding domain (RBD) of MERS-CoV spike protein and DPP4 was determined by crystallography. Animal studies identified rhesus macaques but not hamsters, ferrets, or mice to be susceptible for MERS-CoV. Here, we investigated the role of DPP4 in this observed species tropism. Cell lines of human and nonhuman primate origin were permissive of MERS-CoV, whereas hamster, ferret, or mouse cell lines were not, despite the presence of DPP4. Expression of human DPP4 in nonsusceptible BHK and ferret cells enabled MERS-CoV replication, whereas expression of hamster or ferret DPP4 did not. Modeling the binding energies of MERS-CoV spike protein RBD to DPP4 of human (susceptible) or hamster (nonsusceptible) identified five amino acid residues involved in the DPP4-RBD interaction. Expression of hamster DPP4 containing the five human DPP4 amino acids rendered BHK cells susceptible to MERS-CoV, whereas expression of human DPP4 containing the five hamster DPP4 amino acids did not. Using the same approach, the potential of MERS-CoV to utilize the DPP4s of common Middle Eastern livestock was investigated. Modeling of the DPP4 and MERS-CoV RBD interaction predicted the ability of MERS-CoV to bind the DPP4s of camel, goat, cow, and sheep. Expression of the DPP4s of these species on BHK cells supported MERS-CoV replication. This suggests, together with the abundant DPP4 presence in the respiratory tract, that these species might be able to function as a MERS-CoV intermediate reservoir.
  
  
  OBJECTIVE
  The ongoing outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) has caused 701 laboratory-confirmed cases to date, with 249 fatalities. Although bats and dromedary camels have been identified as potential MERS-CoV hosts, the virus has so far not been isolated from any species other than humans. The inability of MERS-CoV to infect commonly used animal models, such as hamster, mice, and ferrets, indicates the presence of a species barrier. We show that the MERS-CoV receptor DPP4 plays a pivotal role in the observed species tropism of MERS-CoV and subsequently identified the amino acids in DPP4 responsible for this restriction. Using a combined modeling and experimental approach, we predict that, based on the ability of MERS-CoV to utilize the DPP4 of common Middle East livestock species, such as camels, goats, sheep, and cows, these form a potential MERS-CoV intermediate host reservoir species.
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Neeltje
authors.initials: N
authors.lastname: van Doremalen
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Kerri L
authors.initials: KL
authors.lastname: Miazgowicz
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Shauna
authors.initials: S
authors.lastname: Milne-Price
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Trenton
authors.initials: T
authors.lastname: Bushmaker
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Shelly
authors.initials: S
authors.lastname: Robertson
authors.affiliation: Rocky Mountain Veterinary Branch, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Dana
authors.initials: D
authors.lastname: Scott
authors.affiliation: Central Veterinary Research Laboratories, Dubai, Dubai, United Arab Emirates.
authors.forename: Joerg
authors.initials: J
authors.lastname: Kinne
authors.affiliation: Department of Biochemistry, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA.
authors.forename: Jason S
authors.initials: JS
authors.lastname: McLellan
authors.affiliation: Department of Immunology and Microbial Science and Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA vincent.munster@nih.gov.
authors.forename: Vincent J
authors.initials: VJ
authors.lastname: Munster
mesh_terms: D000818:Animals; D002162:Camelus / Q000378:metabolism / Q000821:virology; D002417:Cattle; D002460:Cell Line; D045744:Cell Line, Tumor; D017934:Coronavirus / Q000378:metabolism / Q000472:pathogenicity*; D006224:Cricetinae; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D005289:Ferrets / Q000378:metabolism / Q000821:virology; D006041:Goats / Q000378:metabolism / Q000821:virology; D058507:Host Specificity; D006801:Humans; D058751:Livestock / Q000378:metabolism / Q000821:virology; D008253:Macaca mulatta / Q000378:metabolism / Q000821:virology; D051379:Mice; D008810:Mice, Inbred C57BL; D008877:Middle East; D011323:Primates / Q000378:metabolism / Q000821:virology; D011485:Protein Binding; D000085064:Receptors, Coronavirus; D011991:Receptors, Virus / Q000378:metabolism*; D012136:Respiratory Syncytial Viruses / Q000378:metabolism / Q000472:pathogenicity*; D012756:Sheep / Q000378:metabolism / Q000821:virology; D014709:Vero Cells; D056189:Viral Tropism; D014779:Virus Replication / Q000235:genetics
chemical_list: D000085064:Receptors, Coronavirus; D011991:Receptors, Virus; D018819:Dipeptidyl Peptidase 4