Characterization of the UL16 gene product of herpes simplex virus type 2.
We have raised rabbit polyclonal antisera against a His-tagged herpes simplex virus type 1 (HSV-1) UL16 fusion protein, one of which very specifically reacted with 40 kDa and 41 kDa proteins in the lysates of HSV-1 and HSV-2-infected cells, respectively. Since its reactivity to the 41 kDa protein was clearly eliminated by pre-adsorption with E. coli lysates expressing the UL16 fusion protein, the antiserum was used to characterize the UL16 products of HSV-2. The HSV-2 UL16 protein was produced at the late phase of infection in a manner highly dependent on viral DNA synthesis and was distributed in both the nuclei and the cytoplasma of infected cells. In immunofluorescence studies, the UL16-specific fluorescence in the nuclei was shown to be detected as small discrete granules. On the other hand, the cytoplasmic fluorescence was diffusely distributed around the nucleus at 8 h postinfection but, at later times of infection, it was mainly detected as a mass at a perinuclear region. The analysis on its association with capsids has revealed that the UL16 protein copurified with C capsids but not B and A capsids, and that the association with C capsids was not tight. Moreover, our experiments have shown that a detectable level of the UL16 protein was not associated with extracellular virions, and that the partially purified UL16 proteins had a DNA-binding activity. These observations are consistent with the hypothesis that the UL16 protein plays a role in capsid maturation including DNA packaging/cleavage. We have also determined the complete nucleotide sequence of the HSV-2 UL16 gene and found that a nonstandard initiation codon may be used for its translation.
Oshima
S
S
Laboratory of Virology, Nagoya University School of Medicine, Japan.
Daikoku
T
T
Shibata
S
S
Yamada
H
H
Goshima
F
F
Nishiyama
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral; D000918:Antibody Specificity; D001483:Base Sequence; D002213:Capsid / Q000378:metabolism; D002522:Chlorocebus aethiops; D018387:Codon, Initiator / Q000235:genetics; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D005814:Genes, Viral; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D013347:Subcellular Fractions / Q000378:metabolism / Q000821:virology; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000502:physiology*
D000914:Antibodies, Viral; D018387:Codon, Initiator; D017931:DNA Primers; D014760:Viral Fusion Proteins; D004247:DNA