Nucleolar localization of the UL3 protein of herpes simplex virus type 2.
A rabbit polyclonal antiserum was raised against a recombinant 6 x His-UL3 fusion protein expressed in Escherichia coli and used to examine the intracellular localization of the UL3 protein of herpes simplex virus type 2 (HSV-2). The antiserum reacted specifically with 31 and 34 kDa proteins in HSV-2 186-infected Vero cells and with 31 and 35 kDa proteins in UL3-expressing COS-7 cells. The UL3 protein localized both in the cytoplasm and in five to ten bright fluorescent granules in the nucleus close to the nuclear membrane at 4 h post-infection (p.i.). These structures became bigger at 5 h p.i. and showed doughnut-like forms at 6 h p.i. In transfected Vero cells, the UL3 protein localized exclusively in the nucleoplasm and specifically in the nucleolus. Five deletion mutants of the UL3 protein were constructed for transfection assays and the results showed that the region containing amino acids 100-164 was important for nucleolar localization. Moreover, green fluorescent protein (GFP)-targetting experiments showed that the region containing amino acids 100-164 was able to transport non-nucleolar GFP to the nucleolus as a fusion protein.
Yamada
Hiroshi
H
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-ku, Nagoya 466-8550, Japan1.
Jiang
Yue-Mei
YM
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-ku, Nagoya 466-8550, Japan1.
Zhu
Hong-Yan
HY
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-ku, Nagoya 466-8550, Japan1.
Inagaki-Ohara
Kyoko
K
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-ku, Nagoya 466-8550, Japan1.
Nishiyama
Yukihiro
Y
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-ku, Nagoya 466-8550, Japan1.
D000818:Animals; D000918:Antibody Specificity; D019556:COS Cells; D002466:Cell Nucleolus; D002522:Chlorocebus aethiops; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D018258:Herpesvirus 2, Human / Q000235:genetics; D006801:Humans; D007424:Intracellular Fluid; D008164:Luminescent Proteins / Q000235:genetics; D016296:Mutagenesis; D010750:Phosphoproteins / Q000032:analysis* / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D014709:Vero Cells; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics
D008164:Luminescent Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C082251:UL3 protein, Human herpesvirus 2; D014764:Viral Proteins; D049452:Green Fluorescent Proteins