MOLECULAR BIOLOGY OF HERPES SIMPLEX VIRUSES

单纯疱疹病毒(HSV)是致癌病毒家族的成员， 引起中到重度疾病，特别是在新生儿和 感染或药物导致的免疫缺陷个人，包括 癌症患者，并作为辅助因素与癌症相关 宫颈。此应用程序是OIG的竞争性续订， 整合了四个以分子为中心的长期项目 这些病毒的生物学。目前的研究目标如下： 项目1涉及lambda134.5，这是一种最近发现的病毒基因，其 该产品是中枢神经系统病毒复制所必需的。兰达134.5 负性病毒特异性诱导神经细胞程序性细胞死亡 这就阻止了病毒的繁殖。Lambda134.5 in Transform 有丝分裂前神经母细胞瘤细胞排除凋亡和补体 缺乏这种基因的病毒也会诱导细胞分化。一个 同系物，MyD116，仅与lambda134.5共享63个羧基末端 氨基酸是从髓系白血病细胞中分离出来的。 白细胞介素6.携带MyD116的病毒诱导分化 放置lambda134.5正常复制，不诱导细胞凋亡 神经细胞。我们的目标是确定(I)结构 Lambda134.5和MYD116蛋白的组成，(Ii)其中和lambda134.5 和MyD116蛋白的功能，特别是检测配体和 氨基末端和羧基末端结构域的效应功能 在这些蛋白中，(Iii)可能是lambda134.5和Myd116的受体 效应域和配基结构域以及(Iv)抗细胞凋亡的关系 以及lambda134.5和MyD116的分化功能。此外，它还 注意到在注射&>100,000，LD50剂量的Lambda134.5时 病毒对小鼠是无害的，病毒是能够摧毁的 先前植入中枢神经系统的恶性胶质瘤细胞的致死剂量。 其目的是确定针对中枢神经系统的无毒病毒 癌细胞可以被设计出来。项目2以几个监管机构为中心 核苷化和/或磷酸化的蛋白质以及 最近发现的两种病毒蛋白激酶(UL13和UL13)的底物 美国3)。US3的底物是必需的膜蛋白UL34。在……里面 感染US3减去病毒或感染病毒的细胞 UL34的磷酸化位点发生了突变，UL34蛋白 在免疫沉淀试验中与四种 磷蛋白。UL13的底物是调控的产物 基因阿尔法22。目标是确定(I)功能和 病毒蛋白的鸟苷化和腺化的机制，(Ii) UL13蛋白翻译后修饰的功能，(Iii) 与Ul34蛋白相关的磷酸蛋白四元组的性质和 (4)α22的磷酸化在基因表达中的作用。这个 项目3的目标是解决5‘的功能 Lambda(晚期)病毒未转录和转录的非编码区 基因，特别是确定潜在的链内的作用 根据晚期基因对病毒DNA合成的依赖性进行碱基配对。这个 3个项目非完全依赖于病毒的基因工程。 携带特殊设计的嵌合基因和携带蛋白质 具体修改。

Herpes simplex viruses (HSV), members of a family of oncogenic viruses, cause moderate to severe disease, particularly in neonates and in infection-or drug induced immunologically deficient individuals including cancer patients, and are associated as co-factors with cancer of the cervix. This application is a competing renewal of an OIG which consolidated four long standing projects centering on the molecular biology of these viruses. Current research objectives are as follows: Project 1 concerns lambda134.5, a recently discovered viral gene whose product is essential from viral replication in CNS. The lambda134.5 minus virus induces specifically in neuronal cells programmed cell death which precludes viral multiplication. Lambda134.5 in transformed premitotic neuroblastoma cells precludes apoptosis and complements viruses lacking this gene and also induces cells to differentiate. A homolog, MyD116, sharing with lambda134.5 only the 63 carboxyl terminal amino acids was isolated by others form myelogenous leukemic cells induced to differentiate by interleukin 6. Viruses carrying MyD116 in place lambda134.5 replicate normally and do not induce apoptosis in neuronal cell. Our objectives are to determine (i) the structural components of lambda134.5 and MYD116 proteins, (ii) where and lambda134.5 and MyD116 proteins function and in particular to test for ligand and effector functions of the amino terminal and carboxyl-terminal domains of the proteins, (iii) the receptors of lambda134.5 and Myd116 putative effector and ligand domains and (iv) the relationship of anti-apoptosis and differentiation functions of lambda134.5 and MyD116. In addition it was noted that while injection of > 100,000 LD50 doses of lambda134.5 minus virus is non destructive to mice, the virus is able to destroy lethal doses of malignant glioma cells implanted previously into the CNS. The intent is to determine whether avirulent viruses specific for CNS cancer cells can be designed. Project 2 centers on several regulatory proteins which are nucleotidylated and/or phosphorylated and on the recently identified substrates of two viral protein kinases (UL13 and US3). The substrate of US3 is the essential membrane protein UL34. In cells infected with US3 minus virus or with a virus in which the phosphorylation site in UL34 had been mutagenized, the UL34 protein becomes associated in immune precipitation tests with four phosphoprotein. The substrate of UL13 is the product of the regulatory gene alpha22. The objectives are to determine (i) the function and mechanism of guanylation and adenylation of viral proteins, (ii) the function of post-translation modification of the UL13 protein, (iii) the nature of the phosphoprotein quartet associated with the Ul34 protein and (iv) the role of phosphorylation of alpha22 in gene expression. The objective of Project 3 is to resolve the functions of the 5' untranscribed and transcribed noncoding domains of lambda (late) viral genes and in particular to determine the role of potential intrastrand base pairing on the dependence of late genes on viral DNA synthesis. The 3 projects rely, non exclusively, on the genetic engineering of viruses carrying specifically designed chimeric genes and on proteins carrying specific modifications.