title: The US5 open reading frame of herpes simplex virus type 1 does encode a glycoprotein (gJ).
abstract: Based on sequence analysis, the protein encoded by the US5 open reading frame (ORF) of herpes simplex virus type 1 (HSV-1) was predicted to contain an N-glycosylation site and was given the designation of glycoprotein J (gJ). However, the US5 gene product has not been identified and the identity of gJ as a glycoprotein has not been confirmed. We have cloned and expressed the DNA encoding the complete sequence of the US5 ORF, using a baculovirus expression system. Western blotting, using polyclonal antibody raised against synthetic US5 peptides, revealed two major baculovirus-US5-expressed protein bands with apparent molecular weights of 16-17 and 10 kD. The recombinant US5 was found on the membrane of Spodoptera frugiperda cells and was susceptible to tunicamycin, endoglycosidase H, glycosidase F and partially resistant to endoglycosidase F. Vaccination of mice with baculovirus-expressed US5 did not induce a neutralizing antibody to HSV-1 or provide protection against lethal HSV-1 challenge. However, serum from these vaccinated mice was able to recognize US5 in purified HSV-1 virions by Western blot analyses and on the surface of HSV-1-infected cells by immunofluorescence. These findings establish that US5 does encode a glycoprotein and confirm the appropriateness of naming the US5 gene product gJ.
authors.affiliation: Ophthalmology Research, Cedars-Sinai Medical Center Research Institute, Los Angeles, Calif., 90048, USA. ghiasi@csmc.edu
authors.forename: H
authors.initials: H
authors.lastname: Ghiasi
authors.forename: A B
authors.initials: AB
authors.lastname: Nesburn
authors.forename: S
authors.initials: S
authors.lastname: Cai
authors.forename: S L
authors.initials: SL
authors.lastname: Wechsler
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000821:virology; D003001:Cloning, Molecular; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D017924:Nucleopolyhedroviruses / Q000235:genetics; D016366:Open Reading Frames; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C077970:glycoprotein J, herpesvirus simiae