DEOXYURIDINE METABOLISM IN HERPES LABIALIS

单纯疱疹病毒(HSV)是许多 包括唇疱疹在内的疾病与 单纯疱疹病毒感染与某些肿瘤发生的关系 疾病。单纯疱疹病毒在小鼠体内诱导约50个多肽 被允许感染的细胞，其中一些细胞具有酶 活动。然而，除了HSV诱导的DNA聚合酶 和胸苷激酶，对其他蛋白的作用知之甚少 HSV在复制或抗病毒化疗中诱导的酶。 本研究的目的是确定单纯疱疹病毒是否诱导了 脱氧尿苷三磷酸核苷酸水解酶(dUTPase，EC 2.6.1.23)可用作开发的目标站点 特定的抗病毒化合物，并确定这一作用 酶可能在调节化疗中的作用 特定抗病毒药物的有效性。为了实现这一目标，我们 计划使用纯化的HSV诱导和细胞dUTPase来 阐明有关dUTP酶活性部位的特征，以 确定这些化合物的空间结合性质的差异 并确定这些酶的作用机制(S) 各种汞(II)化合物对dUTP酶有抑制作用。我们 将构建，使用插入和定点突变 DUTPase活性有缺陷的HSV突变体 DUTPase活性改变。这些突变体以及野生型 病毒将用于体内研究，以确定病毒的变化 脱氧尿苷(DURD)池将影响dUTP在 并确定dUTP如何掺入HSV DNA中 DNA影响HSV复制。

Herpes simplex virus (HSV) is the etiological agent of a number of diseases including herpes labialis and there is a correlation between HSV infections and the development of certain neoplastic diseases. HSV induces approximately 50 polypeptides in permissively infected cells and some of these possess enzymatic activity. However, except for the HSV induced DNA polymerase and thymidine kinase, little is known concerning the roles of other HSV-induced enzymes in replication or in antiviral chemotherapy. The goal of this research is to determine whether the HSV induced deoxyuridine triphosphate nucleotidohydrolase (dUTPase, EC 2.6.1.23) can be used as a target site for the development of specific antiviral compounds and to determine what role this enzyme may have in regulating the chemotherapeutic effectiveness of specific antiviral agents. To accomplish this, we plan to use purified HSV induced and cellular dUTPases to elucidate features concerning the active-site of the dUTPases, to determine differences in the steric-binding properties of these enzymes and to determine the mechanism(s) by which these dUTPases are inhibited by various mercury (II) compounds. We will construct, using insertional and site-directed mutagenesis HSV mutants that are defective in dUTPase activity and that have altered dUTPase activity. These mutants as well as wild-type virus will be used in in vivo studies to determine how changes in deoxyuridine (dUrd) pools will effect incorporation of dUTP into HSV DNA and to determine how incorporation of dUTP into the DNA effects HSV replication.