Recently, coronaviruses have generated substantial interest from the biomedical community based on the emergence and isolation of a novel and deadly coronavirus infecting humans. This virus was recently named Middle East Respiratory Syndrome (MERS)-CoV. Like other enveloped viruses, coronaviruses access their host cells by membrane fusion, a process mediated by a specific fusion, or """"""""spike"""""""" protein on the virion. Such viral fusion proteins are often activated by host cell proteases. Mutations in the virus genome can lead to modifications to the proteolytic cleavage sites(s) on the spike that can subsequently regulate membrane fusion, virus entry and host cell tropism. We have identified changes in the MERS-CoV spike protein cleavage site(s) that distinguish it from its closest relatives, as well as most other coronaviruses. These changes allow cleavage-activation of the spike protein by furin, a broadly expressed protease. Furin-mediated activation is unusual in that it occurs during virus entry. The ability of MERS-CoV to utilize furin in this manner may explain the polytropic nature of the virus and its life cycle as a zoonotic coronavirus. In this application, we will characterize the proteolytic cleavage sites of MERS-CoV spike, in comparison to its closest relatives BatCoV- HKU4 and BatCoV-HKU5, and determine the role of furin-mediated cleavage in the context of the polytropic nature of MERS-CoV. More broadly, our goals are to understand how mutations in the CoV spike cleavage sites modulate protease cleavage and fusion activation. Our work will be useful for public health planning and in testing the feasibility of usng furin inhibitors as anti-viral therapeutics for MERS-like CoV infections.

Public Health Relevance

Coronaviruses are currently receiving much attention as agents of infectious disease, due to the emerging outbreak of Middle East respiratory syndrome (MERS). Our studies focus on the ability of the coronaviruses to be activated by host cell proteases prior to membrane fusion with host cells, with changes in protease usage being a major factor in their host/tissue range of the virus. Our studies will serve model for viral pathogenesis and may lead to the development of new antiviral drugs, both for MERS-CoV and for other potential zoonotic coronaviruses.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Exploratory/Developmental Grants (R21)
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Virology - B Study Section (VIRB)
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Park, Eun-Chung
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Cornell University
Schools of Veterinary Medicine
United States
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Millet, Jean Kaoru; Whittaker, Gary R (2018) Physiological and molecular triggers for SARS-CoV membrane fusion and entry into host cells. Virology 517:3-8
Lai, Alex L; Millet, Jean K; Daniel, Susan et al. (2017) The SARS-CoV Fusion Peptide Forms an Extended Bipartite Fusion Platform that Perturbs Membrane Order in a Calcium-Dependent Manner. J Mol Biol 429:3875-3892
White, Judith M; Whittaker, Gary R (2016) Fusion of Enveloped Viruses in Endosomes. Traffic 17:593-614
Millet, Jean K; Séron, Karin; Labitt, Rachael N et al. (2016) Middle East respiratory syndrome coronavirus infection is inhibited by griffithsin. Antiviral Res 133:1-8
Millet, Jean Kaoru; Goldstein, Monty E; Labitt, Rachael N et al. (2016) A camel-derived MERS-CoV with a variant spike protein cleavage site and distinct fusion activation properties. Emerg Microbes Infect 5:e126
Millet, Jean Kaoru; Whittaker, Gary R (2016) Murine Leukemia Virus (MLV)-based Coronavirus Spike-pseudotyped Particle Production and Infection. Bio Protoc 6:
Millet, Jean Kaoru; Whittaker, Gary R (2015) Host cell proteases: Critical determinants of coronavirus tropism and pathogenesis. Virus Res 202:120-34
Millet, Jean Kaoru; Whittaker, Gary R (2014) Host cell entry of Middle East respiratory syndrome coronavirus after two-step, furin-mediated activation of the spike protein. Proc Natl Acad Sci U S A 111:15214-9