Viral and cellular gene regulation during lytic KSHV replication

这一应用的广泛、长期的目标是了解卡波西的调节蛋白是如何 肉瘤相关疱疹病毒(KSHV/HHV-8)在裂解过程中控制病毒和细胞基因的表达 复制。裂解基因产物可能在KSHV的发病机制中起作用，有助于 KSHV感染细胞和邻近细胞的增殖、存活和避免免疫反应的能力。 因此，了解裂解基因表达的调控机制与KSHV- 相关的发病机制仍然是发病率和死亡率的一个重要原因，特别是在 艾滋病毒和KSHV感染率高的地区。 KSHV的ORF57蛋白在KSHV复制早期表达，是KSHV的一个家族成员。 疱疹病毒蛋白，在疱疹病毒复制中起重要作用。ORF57具有独特的监管 性质，转录后增强许多无内含子基因的表达。此外，ORF57 增强特定细胞基因的表达。ORF57很可能通过物理绑定来发挥其许多影响 并调节其稳定性和核输出。关于这种机制仍有许多未知之处。 哪个ORF57影响RNA的加工以及它的作用特异性是如何被确定的。 因此，我们提出了三个综合目标来研究ORF57在KSHV生物学中的作用。在第一个 目的，我们将确定ORF57与mRNA结合的机制以及ORF57的特异性是如何确定的。 在第二个目标中，我们将确定对ORF57功能重要的主要细胞蛋白， 特别是在核出口和信使核糖核酸加工方面。在第三个目标中，我们将确定哪个目标 ORF57的功能对于利用分子遗传学复制KSHV是必不可少的。我们还将确定 ORF57对血管内皮细胞和B淋巴细胞基因表达的特异性影响 KSHV是一种与癌症的发展有关的病毒，特别是在那些身体虚弱的人身上 免疫系统，例如那些感染了艾滋病毒的人。研究病毒是如何繁殖的对 了解它是如何影响它感染的细胞并将它们转化为肿瘤细胞的。这样的研究也有 有可能确定针对病毒的新药的靶点，这些新药可能有助于预防或治疗感染。

The broad, long term objective of this application is to understand how regulatory proteins of Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) control viral and cellular gene expression during lytic replication. Lytic gene products are likely to have a role in pathogenesis of KSHV, contributing to the ability of KSHV-infected cells and adjacent cells to proliferate, survive and avoid immune responses. Understanding the mechanisms by which lytic gene expression is regulated is therefore relevant to KSHV- related pathogenesis which continues to be a significant cause of morbidity and mortality, particularly in areas with high endemic rates of HIV and KSHV infection. The ORF57 protein of KSHV is expressed early during KSHV replication and is a member of a family of herpesvirus proteins that plays an essential role in herpesvirus replication. ORF57 has unique regulatory properties, post-transcriptionally enhancing expression of many intronless genes. In addition, ORF57 enhances expression of specific cell genes. ORF57 is likely to exert many of its effects by physically binding to mRNA and modulating its stability and nuclear export. Much remains unknown about the mechanismsby which ORF57 affects RNA processing and how its specificity of action is determined. We therefore propose three integrated aims to investigate the role of ORF57 in KSHV biology. In the first aim, we will define the mechanisms by which ORF57 binds mRNA and how ORF57 specificity is determined. In the second aim, we will identify the major cellular proteins that are important for ORF57 function, especially with regard to nuclear export and mRNA processing. In the third aim, we will determine which functions of ORF57 are essential for KSHV replication using molecular genetics. We will also determine the specific effects of ORF57 on gene expression in endothelial cells and B lymphocytes. KSHV is a virus that is linked to the development of cancer, particularly in those people who have weakened immune systems, such as those with HIV infection. Studying how the virus reproduces is important to understand how it affects the cells it infects and converts them to tumor cells. Such studies also have the potential to identify targets for new drugs against the virus that may help prevent or treat infection.