Murata Takayuki, Goshima Fumi, Nishizawa Yuji, Daikoku Tohru, Takakuwa Hiroki, Ohtsuka Kenzo, Yoshikawa Tetsushi, Nishiyama Yukihiro
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Nagoya, Aichi 466-8550, Japan.
Microbiol Immunol. 2002;46(10):707-19. doi: 10.1111/j.1348-0421.2002.tb02755.x.
We previously reported the establishment of an HEp2 cell line which expresses the US3 protein kinase (PK) of herpes simplex virus type 2 (HSV-2) upon induction with IPTG. Here we report that expression, phosphorylation and ubiquitination of cytokeratin 17 (CK17) are enhanced in US3-expressing HEp2 cells. In vitro kinase and co-immunoprecipitation assays provided evidence that US3 PK directly phosphorylates CK17. Expression of US3 PK caused a significant decrease in filamentous staining of CK17, suggesting that phosphorylation of CK17 by US3 PK causes a disruption of intermediate filaments. Our observations suggest a role for US3 in the regulation of CKs and intermediate filaments in cells. Moreover, we found that infection of a keratinocyte-derived cell line, A431, with a US3-deficient virus, results in cytopathic effects that are morphologically distinct from those induced by wild-type and revertant viruses, suggesting that US3 PK may be important for interaction between HSV-2 and peripheral epithelial cells.
我们之前报道过建立了一种HEp2细胞系，在用异丙基-β-D-硫代半乳糖苷（IPTG）诱导时可表达2型单纯疱疹病毒（HSV-2）的US3蛋白激酶（PK）。在此我们报道，在表达US3的HEp2细胞中，细胞角蛋白17（CK17）的表达、磷酸化和泛素化增强。体外激酶和免疫共沉淀试验提供了证据，证明US3 PK可直接使CK17磷酸化。US3 PK的表达导致CK17的丝状染色显著减少，这表明US3 PK对CK17的磷酸化会导致中间丝的破坏。我们的观察结果表明，US3在细胞中细胞角蛋白（CKs）和中间丝的调节中发挥作用。此外，我们发现，用一种缺失US3的病毒感染角质形成细胞来源的细胞系A431，会导致细胞病变效应，其形态与野生型和回复病毒诱导的不同，这表明US3 PK可能对HSV-2与外周上皮细胞之间的相互作用很重要。
