title: Vaccinia virus encodes I5, a small hydrophobic virion membrane protein that enhances replication and virulence in mice.
abstract: The vaccinia virus I5L open reading frame encodes a 79-amino-acid protein, with two predicted transmembrane domains, that is conserved among all sequenced members of the chordopoxvirus subfamily. No nonpoxvirus homologs or functional motifs have been recognized, and the role of the I5 protein remains unknown. We found that synthesis of I5 was dependent on viral DNA replication and occurred exclusively at late times, consistent with a consensus late promoter motif adjacent to the start of the open reading frame. I5 was present in preparations of purified virions and could be extracted with nonionic detergent, suggesting membrane insertion. Transmission electron microscopy of immunogold-labeled thawed cryosections of infected cells revealed the association of an epitope-tagged I5 with the membranes of immature and mature virions. Viable I5L deletion and frameshift mutants were constructed and found to replicate like wild-type virus in a variety of cell lines and primary human epidermal keratinocytes, indicating that the protein was dispensable for in vitro cultivation. However, mouse intranasal challenge experiments indicated that a mutant virus with a frameshift resulting in a stop codon near the N terminus of I5 was attenuated compared to control virus. The attenuation was correlated with clearance of mutant viruses from the respiratory tract and with less progression and earlier resolution of pathological changes. We suggest that I5 is involved in an aspect of host defense that is evolutionarily conserved although a role in cell tropism should also be considered.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 33 North Drive, MSC 3210, Bethesda, MD 20892-3210, USA.
authors.forename: Cindy L
authors.initials: CL
authors.lastname: Sood
authors.forename: Jerrold M
authors.initials: JM
authors.lastname: Ward
authors.forename: Bernard
authors.initials: B
authors.lastname: Moss
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D005260:Female; D015967:Gene Expression Regulation, Viral; D006801:Humans; D008168:Lung / Q000166:cytology / Q000473:pathology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009297:Nasal Mucosa / Q000166:cytology / Q000473:pathology / Q000821:virology; D016366:Open Reading Frames; D016415:Sequence Alignment; D018523:Tropism; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D014759:Viral Envelope Proteins