title: A Molecular Sensor To Characterize Arenavirus Envelope Glycoprotein Cleavage by Subtilisin Kexin Isozyme 1/Site 1 Protease.
abstract: Arenaviruses are emerging viruses including several causative agents of severe hemorrhagic fevers in humans. The advent of next-generation sequencing technology has greatly accelerated the discovery of novel arenavirus species. However, for many of these viruses, only genetic information is available, and their zoonotic disease potential remains unknown. During the arenavirus life cycle, processing of the viral envelope glycoprotein precursor (GPC) by the cellular subtilisin kexin isozyme 1 (SKI-1)/site 1 protease (S1P) is crucial for productive infection. The ability of newly emerging arenaviruses to hijack human SKI-1/S1P appears, therefore, to be a requirement for efficient zoonotic transmission and human disease potential. Here we implement a newly developed cell-based molecular sensor for SKI-1/S1P to characterize the processing of arenavirus GPC-derived target sequences by human SKI-1/S1P in a quantitative manner. We show that only nine amino acids flanking the putative cleavage site are necessary and sufficient to accurately recapitulate the efficiency and subcellular location of arenavirus GPC processing. In a proof of concept, our sensor correctly predicts efficient processing of the GPC of the newly emergent pathogenic Lujo virus by human SKI-1/S1P and defines the exact cleavage site. Lastly, we employed our sensor to show efficient GPC processing of a panel of pathogenic and nonpathogenic New World arenaviruses, suggesting that GPC cleavage represents no barrier for zoonotic transmission of these pathogens. Our SKI-1/S1P sensor thus represents a rapid and robust test system for assessment of the processing of putative cleavage sites derived from the GPCs of newly discovered arenavirus by the SKI-1/S1P of humans or any other species, based solely on sequence information.
  
  
  OBJECTIVE
  Arenaviruses are important emerging human pathogens that can cause severe hemorrhagic fevers with high mortality in humans. A crucial step in productive arenavirus infection of human cells is the processing of the viral envelope glycoprotein by the cellular subtilisin kexin isozyme 1 (SKI-1)/site 1 protease (S1P). In order to break the species barrier during zoonotic transmission and cause severe disease in humans, newly emerging arenaviruses must be able to hijack human SKI-1/S1P efficiently. Here we implement a newly developed cell-based molecular sensor for human SKI-1/S1P to characterize the processing of arenavirus glycoproteins in a quantitative manner. We further use our sensor to correctly predict efficient processing of the glycoprotein of the newly emergent pathogenic Lujo virus by human SKI-1/S1P. Our sensor thus represents a rapid and robust test system with which to assess whether the glycoprotein of any newly emerging arenavirus can be efficiently processed by human SKI-1/S1P, based solely on sequence information.
authors.affiliation: Institute of Microbiology, University Hospital Center and University of Lausanne, Lausanne, Switzerland.
authors.forename: Joel
authors.initials: J
authors.lastname: Oppliger
authors.affiliation: Institute of Microbiology, University Hospital Center and University of Lausanne, Lausanne, Switzerland.
authors.forename: Joel Ramos
authors.initials: JR
authors.lastname: da Palma
authors.affiliation: Institute of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Dominique J
authors.initials: DJ
authors.lastname: Burri
authors.affiliation: Viral Special Pathogens Branch, U.S. Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Eric
authors.identifier: http://orcid.org/0000-0003-3398-8628
authors.initials: E
authors.lastname: Bergeron
authors.affiliation: Université Bordeaux, LAMC, Pessac, France INSERM, UMR 1029, Pessac, France.
authors.forename: Abdel-Majid
authors.initials: AM
authors.lastname: Khatib
authors.affiliation: Viral Special Pathogens Branch, U.S. Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Christina F
authors.initials: CF
authors.lastname: Spiropoulou
authors.affiliation: Institute of Microbiology, University Hospital Center and University of Lausanne, Lausanne, Switzerland Antonella.Pasquato@chuv.ch Stefan.Kunz@chuv.ch.
authors.forename: Antonella
authors.initials: A
authors.lastname: Pasquato
authors.affiliation: Institute of Microbiology, University Hospital Center and University of Lausanne, Lausanne, Switzerland Antonella.Pasquato@chuv.ch Stefan.Kunz@chuv.ch.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kunz
mesh_terms: D000818:Animals; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D063927:Lujo virus / Q000502:physiology*; D015336:Molecular Probe Techniques; D043484:Proprotein Convertases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012697:Serine Endopeptidases / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D014759:Viral Envelope Proteins; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; C115575:membrane-bound transcription factor peptidase, site 1