title: A viral member of the ERV1/ALR protein family participates in a cytoplasmic pathway of disulfide bond formation.
abstract: Proteins of the ERV1/ALR family are encoded by all eukaryotes and cytoplasmic DNA viruses for which substantial sequence information is available. Nevertheless, the roles of these proteins are imprecisely known. Multiple alignments of ERV1/ALR proteins indicated an invariant C-X-X-C motif, but no similarity to the thioredoxin fold was revealed by secondary structure predictions. We chose a virus model to investigate the role of these proteins as thiol oxidoreductases. When cells were infected with a mutant vaccinia virus in which the E10R gene encoding an ERV1/ALR family protein was repressed, the disulfide bonds of three other viral proteins-namely, the L1R and F9L proteins and the G4L glutaredoxin-were completely reduced. The same outcome occurred when Cys-43 or Cys-46, the putative redox cysteines of the E10R protein, was mutated to serine. These two cysteines were disulfide bonded during a normal virus infection but not if the synthesis of other viral late proteins was inhibited or the E10R protein was expressed by itself in uninfected cells, suggesting a requirement for an upstream viral thiol oxidoreductase. Remarkably, the cysteine-containing domains of the E10R and L1R viral membrane proteins and the glutaredoxin are in the cytoplasm, in which assembly of vaccinia virions occurs, rather than in the oxidizing environment of the endoplasmic reticulum. These data indicated a viral pathway of disulfide bond formation in which the E10R protein has a central role. By extension, the ERV1/ALR family may represent a ubiquitous class of cellular thiol oxidoreductases that interact with glutaredoxins or thioredoxins.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: T G
authors.initials: TG
authors.lastname: Senkevich
authors.forename: C L
authors.initials: CL
authors.lastname: White
authors.forename: E V
authors.initials: EV
authors.lastname: Koonin
authors.forename: B
authors.initials: B
authors.lastname: Moss
mesh_terms: D000595:Amino Acid Sequence; D003545:Cysteine / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D016366:Open Reading Frames; D050862:Oxidoreductases Acting on Sulfur Group Donors; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D014616:Vaccinia virus / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D004220:Disulfides; D005656:Fungal Proteins; C120898:KMT2D protein, human; D024101:Mitochondrial Proteins; D009363:Neoplasm Proteins; D029701:Saccharomyces cerevisiae Proteins; D050862:Oxidoreductases Acting on Sulfur Group Donors; C073790:ERV1 protein, S cerevisiae; D003545:Cysteine