Definition of structural organization and enzymology of the EBV protein kinase.

描述（由申请人提供）：疱疹病毒编码的蛋白激酶（HPKs）最近引起了越来越多的关注，因为越来越多的证据表明其参与病毒生命周期的不同方面，使其成为抗病毒治疗的吸引人的靶点。该提案的广泛目标是研究和开发针对人类病原体Epstein-Barr病毒（EBV）的新型安全有效的抗病毒剂的病毒编码靶标。该项目采用综合方法：将通过分子生物学和生物化学技术研究EBV编码的蛋白激酶（EBV-PK）的结构和性质，并探索基于小分子和肽的抑制剂策略。第一个具体目标是通过诱变研究EBV-PK结构，表征其对蛋白质底物和更昔洛韦（GCV）的活性，并开发用于快速有效筛选抑制剂的测定法。将在体外和细胞培养物中研究EBV-PK突变体的活性。第二个具体目标是研究小分子和肽类抑制剂对激酶的抑制作用。将在细胞培养物中进一步测试选定的EBV-PK抑制剂抑制病毒复制的能力。因此，本申请旨在表征对EBV复制至关重要的独特激酶，并鉴定将有效抑制该酶的化合物，从而潜在地抑制病毒。因此，它解决了一般的生物学问题和药学问题。鉴于与EBV感染相关的疾病越来越多，其中许多是众所周知的难以治疗，EBV靶向治疗方法的开发和测试是必要的，以试图设计更好的治疗方法。该提案中概述的研究将促进对EBV生物学的理解，并评估开发有效治疗EBV相关疾病的新靶点。

DESCRIPTION (provided by applicant): Herpesvirus-encoded protein kinases (HPKs) have attracted increasing attention lately due to the growing evidence of their involvement in different aspects of the viral life cycle, making them appealing targets for antiviral therapy. The broad objectives of this proposal are to study and exploit a virally encoded target for novel safe and effective antiviral agents against the human pathogen Epstein-Barr virus (EBV). The project takes a combined approach: the structure and properties of the EBV-encoded protein kinase (EBV-PK) will be studied by molecular biology and biochemistry techniques, and in search for the inhibitors both small-molecule and peptide-based strategies will be explored. The first specific aim is to study EBV-PK structure by mutagenesis, characterize its activity in regard to protein substrates and ganciclovir (GCV) and develop an assay for fast and effective screening of inhibitors. The activity of EBV-PK mutants will be studied both in vitro and in cell culture. The second specific aim is designed to study inhibition of the kinase by small-molecule and peptide-based inhibitors. Selected EBV-PK inhibitors will be further tested in cell culture for their ability to inhibit viral replication. Thus, this application aims to characterize a unique kinase, which is crucial for EBV replication, and to identify compounds that will efficiently inhibit this enzyme, potentially inhibiting the virus. Therefore it addresses a general biological question and a pharmaceutical problem. PUBLIC HEALTH RELEVANCE Given the growing number of conditions associated with EBV infection, many of which are notoriously difficult to treat, the development and testing of EBV-targeting therapy approaches is warranted in attempts to devise better treatments. The studies outlined in this proposal will advance understanding of EBV biology and evaluate a novel target for the development of the effective treatment of EBV-associated diseases.