Reddy S M, Cox E, Iofin I, Soong W, Cohen J I
Medical Virology Section, Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892, USA.
J Virol. 1998 Oct;72(10):8083-8. doi: 10.1128/JVI.72.10.8083-8088.1998.
Varicella-zoster virus (VZV) encodes five gene products that do not have homologs in herpes simplex virus. One of these genes, VZV open reading frame 32 (ORF32), is predicted to encode a protein of 16 kDa. VZV ORF32 protein was shown to be phosphorylated and located in the cytosol of virus-infected cells. Antibody to ORF32 protein immunoprecipitated 16- and 18-kDa phosphoproteins from VZV-infected cells. Since VZV encodes two protein kinases that might phosphorylate ORF32 protein, immunoprecipitations were performed with cells infected with VZV mutants unable to express either of the viral protein kinases. Cells infected with VZV unable to express the ORF66 protein kinase contained both the 16- and 18-kDa ORF32 phosphoproteins; however, cells infected with the VZV ORF47 protein kinase mutant showed only the 16-kDa ORF32 phosphoprotein. Treatment of [35S]methionine-labeled proteins with calf intestine alkaline phosphatase resulted in a decrease in size of the ORF32 proteins from 16 and 18 kDa to 15 and 17 kDa, respectively. VZV unable to express ORF32 protein replicated in human melanoma cells to titers similar to those seen with parental virus; however, VZV unable to express ORF32 was impaired for replication in U20S osteosarcoma cells. Thus, VZV ORF32 protein is posttranslationally modified by the ORF47 protein kinase. Since the VZV ORF47 protein kinase has recently been shown to be critical for replication in human fetal skin and lymphocytes, its ability to modify the ORF32 protein suggests that the latter protein may have a role for VZV replication in human tissues.
水痘带状疱疹病毒(VZV)编码五种在单纯疱疹病毒中没有同源物的基因产物。其中一个基因,VZV开放阅读框32(ORF32),预计编码一种16 kDa的蛋白质。VZV ORF32蛋白被证明会发生磷酸化,并位于病毒感染细胞的胞质溶胶中。针对ORF32蛋白的抗体从VZV感染细胞中免疫沉淀出16 kDa和18 kDa的磷酸化蛋白。由于VZV编码两种可能使ORF32蛋白磷酸化的蛋白激酶,因此对感染了无法表达任何一种病毒蛋白激酶的VZV突变体的细胞进行了免疫沉淀。感染了无法表达ORF66蛋白激酶的VZV的细胞中同时含有16 kDa和18 kDa的ORF32磷酸化蛋白;然而,感染VZV ORF47蛋白激酶突变体的细胞仅显示出16 kDa的ORF32磷酸化蛋白。用小牛肠碱性磷酸酶处理[35S]甲硫氨酸标记的蛋白后,ORF32蛋白的大小分别从16 kDa和18 kDa降至15 kDa和17 kDa。无法表达ORF32蛋白的VZV在人黑色素瘤细胞中复制,其滴度与亲本病毒相似;然而,无法表达ORF32的VZV在U20S骨肉瘤细胞中的复制受到损害。因此,VZV ORF32蛋白在翻译后被ORF47蛋白激酶修饰。由于最近已证明VZV ORF47蛋白激酶对在人胎儿皮肤和淋巴细胞中的复制至关重要,其修饰ORF32蛋白的能力表明后者蛋白可能在VZV在人体组织中的复制中起作用。