Biosynthesis of Adenovirus Early RNA's

这项研究的长期目标是了解腺病毒E1 A蛋白是如何激活 从病毒启动子转录，E1 A如何修饰细胞基因的转录， 增殖，腺病毒E1 B-55 K蛋白如何抑制p53功能，以及如何识别人类肿瘤 通过瘤内注射E1 B-55 K缺失突变体可以成功治疗。控制基因 表达是几乎所有生物过程的基本方面;基因表达的异常控制 有助于许多病理过程，如癌症的发展。了解 详细的转录控制的分子机制应该允许设计治疗干预， 治疗人类疾病，如癌症。我们将使用显微荧光方法来分析相互作用 E1 A保守区3和其他激活结构域与介体复合物，一般转录 因子和活细胞中的其他共激活剂复合物。我们将使用染色质免疫沉淀（ChIP）， 分析E1 A对病毒染色质上PIC组装和Pol II重起始的影响。我们将 使用ChIP芯片检测来确定E1 A N-末端区域与CBP和p300的结合如何导致 在最近发现的组蛋白H3 K18乙酰化的重新分布中， 转录激活我们将分析E1 B-55 K通过在细胞中束缚p53来抑制p53功能的能力。 PML核小体依赖于E1 B-55 K类小泛素化，并完全抑制p53激活功能 通过由新发现的E1 B-55 K SUM 01-p53连接酶活性诱导的SUM 01修饰。我们还将 研究E1 B-55 K通过阻止抗病毒反应刺激病毒晚期基因表达的模型 抑制病毒mRNA核输出和翻译。我们将继续研究目前的结果， 这种抗病毒应答的诱导需要由线性病毒激活的MRN复合物激活ATM DNA.这项工作应该会导致对分子相互作用的更全面的理解， 人类细胞中的转录。了解E1 B-55 K刺激病毒复制的机制 不同的人类肿瘤细胞系应该允许临床医生确定哪些人类肿瘤可以被治疗 通过瘤内注射E1 B-55 K缺失突变体有效地治疗。

The long term goals of the proposed research are to understand how the adenovirus E1A protein activates transcription from viral promoters, how E1A modifies transcription of cellular genes that induce cell proliferation, how adenovirus E1B-55K protein inhibits p53 function, and how human tumors can be identified that might be successfully treated by intratumoral injection of an E1B-55K deletion mutant. Control of gene expression is a fundamental aspect of nearly all biological processes; abnormal contol of gene expression contributes to many pathological processes such as the development of cancer. Understanding the molecular mechanisms of transcription control in detail should allow the design of therapeutic interventions to treat human disease such as cancer. We will use microscopic fluorescent methods to analyze the interaction of E1A conserved region 3 and other activation domains with mediator complexes, general transcription factors, and other co-activator complexes in living cells. We will use chromatin-immunoprecipitation (ChIP) to analyze the influence of E1A on PIC assembly and re-initiation by Pol II on viral chromatin in vivo. We will use ChIP on chip assays to determine how the binding of the E1A N-terminal region to CBP and p300 results in the recently discovered re-distribution of histone H3K18 acetylation, a modification that correlates with transcriptional activation. We will analyze the ability of E1B-55K to inhibit p53 function by tethering p53 in PML nuclear bodies dependent on E1B-55K sumoylation, and complete inhibition of p53 activation function by SUMO1-modification induced by a newly discovered E1B-55K SUM01-p53 ligase activity. We will also pursue the model that E1B-55K stimulates viral late gene expression by preventing an anti-viral response that inhibits viral mRNA nuclear export and translation. We will pursue current results suggesting that induction of this anti-viral response requires activation of ATM by MRN complexes activated by linear viral DNA. This work should lead to a more complete understanding of molecular interactionsthat control transcription in human cells. Understanding the mechanism by which E1B-55K stimulates viral replication in various human tumor cell lines should allow clinicians to determine which human tumors might betreated effectively by intratumoral injection of an E1B-55K deletion mutant.