Identification and characterization of the UL7 gene product of herpes simplex virus type 2.
We have raised a rabbit polyclonal antiserum against a recombinant 6x His-tagged herpes simplex virus type 2 (HSV-2) UL7 fusion protein expressed in Escherichia coli. The antiserum specifically reacted with a 33 kDa protein in HSV-1 and HSV-2-infected cell lysates, and was used to characterize the UL7 gene product of HSV-2. The UL7 protein was produced in the late phase of infection, and its synthesis was highly inhibited, but not abolished by the addition of acyclovir (ACV). The UL7 protein associated with extracellular virions and also with all types of capsids, including A, B, and C capsids, though the association seemed to be weak. Indirect immunofluorescence studies revealed that at 9 h postinfection, UL7 specific fluorescence was detected in part or all of the nucleus, and the specific fluorescence colocalized with the scaffold protein ICP35. However, at later times postinfection, the UL7 protein was mainly detected as a mass in a juxtanuclear cytoplasmic region. In addition, transmission immunoelectron microscopy (TIEM) confirmed the association of the UL7 protein with intracellular capsids and virions in HSV-2-infected cells. The HSV-2 UL7 protein contained a domain highly conserved in all herpesviruses, part of which exhibited a homology with domains in the fission yeast Schizosaccharomyces pombe DNA topoisomerase III. We discuss the possibility that the UL7 protein may play a supplementary role in the viral DNA cleavage/packaging process.
Nozawa
Naoki
N
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Japan.
Daikoku
Tohru
T
Yamauchi
Yohei
Y
Takakuwa
Hiroki
H
Goshima
Fumi
F
Yoshikawa
Tetsushi
T
Nishiyama
Yukihiro
Y
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000918:Antibody Specificity; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D005814:Genes, Viral; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D020539:Sequence Analysis, Protein; D014709:Vero Cells; D014771:Virion / Q000378:metabolism
D000914:Antibodies, Viral; D036022:Capsid Proteins