Studies of latent infection of herpes simplex virus and protein kinase

蛋白激酶(PKs)在真核细胞中具有广泛的功能。这些包括新陈代谢、细胞周期、激素反应以及转录和翻译的控制。最常被研究的真核蛋白是那些修饰丝氨酸和苏氨酸残基的蛋白，以及那些修饰酪氨酸残基的蛋白。已有研究表明，单纯疱疹病毒1型(HSV-1)的许多蛋白是磷酸化的，HSV-1编码病毒蛋白，但病毒蛋白的磷酸化在HSV-1感染中的作用尚不清楚。本研究探讨了蛋白酪氨酸激酶(PTK)抑制剂金雀异黄素和酪氨酸蛋白以及蛋白激酶C(PKC)抑制剂1-(5-异喹啉-磺酰基)-2-甲基哌嗪(H-7)对HSV-1复制和病毒蛋白磷酸化的影响。当染料木素浓度大于25µM时，抑制蛋白酪氨酸激酶的染料木素和李素抑制单纯疱疹病毒1型的复制，而相关的复合…更多不抑制PTK的ONE不影响HSV-1的复制。PTKs的合成抑制剂Tyrphostin 9、47、B42和B46均能抑制HSV-1的空斑形成，但Tyrphostin 9、B42和B46的抗病毒作用更具选择性。感染细胞多肽(ICP6、17/18、19/20、25)为酪氨酸磷酸化蛋白。具有酪氨酸残基磷酸化和酪蛋白9抑制作用的HSV-1蛋白是UL47基因的产物，即被膜蛋白VP13/14。PKC抑制剂H-7也抑制HSV-1的复制。另一方面，PKC激活剂12-O-十四酰佛波醇-13-乙酸酯促进了HSV-1诱导的A431表皮样癌细胞合体的形成，提示PKC的激活促进了疱疹粘液皮损中多核巨细胞的形成。本项目还建立了口腔和唾液腺肿瘤的动物模型。未来，我们将测试上述PK抑制剂对HSV-1和体内肿瘤形成的抑制作用。较少

Protein kinases (PKs) are involved in a wide variety of functions in eukaryotic cells. These include metabolism, cell cycle, hormone response, and control of transcription and translation. The most commonly studied eukaryotic PKs are those which modify serine and threonine residues and those which modify tyrosine residues. It has been shown that a number of proteins of herpes simplex virus type 1 (HSV- 1) are phosphorylated and HSV-1 encodes viral PKs, but the role of phosphorylation of viral proteins in HSV-1 infection has not been clarified. In the present study, the effects of PK inhibitors including protein tyrosine kinase (PTK) inhibitors, genistein and tyrphostin, and protein kinase C (PKC) inhibitor, 1-(5-isoquinoline- sulfonyl)-2-methylpiperazine (H-7), on the replication of HSV-1 and phosphorylation of viral proteins were examined. Genistein and prunetin which inhibit PTK inhibited the replication of HSV-1 at genistein concentrations of more than 25muM,whereas the related comp … More ounds, which do not inhibit PTK,did not affect the replication of HSV-1. Tyrphostins 9,47, B42, and B46, synthetic inhibitors of PTKs, all inhibited the plaque formation of HSV-1, but Tyrphostin 9, B42 and B46 exhibited more selective antiviral effect. Infected cell polypeptides (ICP) 6,17/18,19/20, and 25 were found to be tyrosine-phosphorylated proteins. The HSV-1 proteins which have phosphorylated tyrosine residues and suppressed by tyrphostin 9 most significantly were the products of the UL47 gene, the tegument proteins VP13/14. PKC inhibitor H-7 also inhibited the replication of HSV-1. On the other hand, PKC activator 12-o-teradecanoyl phorbol-13-acetate enhanced the HSV-1-induced syncytium formation in A431 epidermoid carcinoma cells, suggesting that PKC activation promotes the development of multinucleated giant cells in herpetic muco-cutaneous lesions. In this project, animal models of oral and salivary gland tumors were also established. In future, we will test the inhibitory effect of above described PK inhibitors on both HSV-1 and tumor formation in vivo. Less