title: Species-specific regions of occludin required by hepatitis C virus for cell entry.
abstract: Hepatitis C virus (HCV) is a leading cause of liver disease worldwide. As HCV infects only human and chimpanzee cells, antiviral therapy and vaccine development have been hampered by the lack of a convenient small-animal model. In this study we further investigate how the species tropism of HCV is modulated at the level of cell entry. It has been previously determined that the tight junction protein occludin (OCLN) is essential for HCV host cell entry and that human OCLN is more efficient than the mouse ortholog at mediating HCV cell entry. To further investigate the relationship between OCLN sequence and HCV species tropism, we compared OCLN proteins from a range of species for their ability to mediate infection of naturally OCLN-deficient 786-O cells with lentiviral pseudoparticles bearing the HCV glycoproteins. While primate sequences function equivalently to human OCLN, canine, hamster, and rat OCLN had intermediate activities, and guinea pig OCLN was completely nonfunctional. Through analysis of chimeras between these OCLN proteins and alanine scanning mutagenesis of the extracellular domains of OCLN, we identified the second half of the second extracellular loop (EC2) and specific amino acids within this domain to be critical for modulating the HCV cell entry factor activity of this protein. Furthermore, this critical region of EC2 is flanked by two conserved cysteine residues that are essential for HCV cell entry, suggesting that a subdomain of EC2 may be defined by a disulfide bond.
authors.affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York, NY, USA.
authors.forename: Maria L
authors.initials: ML
authors.lastname: Michta
authors.forename: Sharon E
authors.initials: SE
authors.lastname: Hopcraft
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Narbus
authors.forename: Zerina
authors.initials: Z
authors.lastname: Kratovac
authors.forename: Benjamin
authors.initials: B
authors.lastname: Israelow
authors.forename: Marion
authors.initials: M
authors.lastname: Sourisseau
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Evans
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004285:Dogs; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D006526:Hepatitis C / Q000235:genetics / Q000378:metabolism* / Q000662:veterinary* / Q000821:virology; D006524:Hepatitis, Viral, Animal / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D062793:Occludin; D011323:Primates; D051381:Rats; D016415:Sequence Alignment; D013045:Species Specificity; D053586:Virus Internalization
chemical_list: D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; C568986:Ocln protein, mouse; C568987:Ocln protein, rat