MOLECULAR BIOLOGY OF RUBELLA VIRUS

风疹病毒的分子生物学

• 批准号: 2442417
• 负责人: TERYL K FREY
• 金额: $ 17.71万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2442417
• 关键词: RNA virus; Rubivirus; active immunization; capsid; complementary DNA; endopeptidases; genetic promoter element; genome; hepatitis virus; laboratory mouse; laboratory rabbit; molecular chaperones; nucleic acid sequence; open reading frames; plasmids; posttranslational modifications; protein structure; site directed mutagenesis; transfection /expression vector; vaccine development; vector vaccine; viral vaccines; virion; virus RNA; virus genetics; virus protein; virus replication

Rubella virus (RUB) is a major human pathogen. RUB infection during pregnancy can lead to severe birth defects (congenital rubella syndrome or CRS). Rubella is controlled in the U.S. by use of live, attenuated vaccines. However, two challenges confront the vaccination program. First, the virus is endemic in much of the world, necessitating a continuing vigilant vaccination effort to prevent reintroduction as occurred between 1989 and 1991. Secondly, in adult women, the vaccine has been associated with chronic arthritis thought to be due to the persistence of the vaccine virus. Another important medical aspect of RUB is that CRS patients suffer from a high incidence of diabetes, providing the best association of a specific human virus with an autoimmune disease. The long-term goal of the proposed research is a thorough study of the molecular biology of RUB. Despite its medical importance, progress on the molecular biology of RUB has been slow, primarily because RUB replicates slowly and to low titers in cell culture. A recent landmark event in the molecular biology of RUB was development of an "infectious clone". An infectious clone is a complete cDNA copy of an RNA virus genome contained in a plasmid from which infectious RNA transcripts can be synthesized in vitro. The first specific aim of the proposed research is to use the infectious clone for two ends: 1). to study by site-directed mutagenesis the function of specific regions of the genome and the encoded proteins, and 2). to develop an infectious clone based on the live, attenuated vaccine virus. The vaccine infectious clone will be introduced into a "genetic immunization" vector, a plasmid from which the genome RNA will be expressed in cells transfected with the recombinant plasmid alone, initiating virus replication. This recombinant vector could be the basis of a DNA vaccine which could be used in worldwide vaccination programs against RUB. The vaccine infectious clone could also be used to modify the vaccine to avert the complications encountered in adult women. The second specific aim is to study the protease responsible for processing the virus replicase or nonstructural proteins (NSPs). As part of this effort, the NSP open reading frame (ORF1) of hepatitis E virus (HEV) will be expressed to analyze its processing. Computer alignment has revealed that the NSP-ORFs of RUB and HEV are more closely related to each other than to other viruses, despite the disparity in structure of RUB and HEV. HEV is the major cause of enterically transmitted non-A, non-B hepatitis and is associated with a high mortality rate in pregnant women. Study of the NSP proteases of RUB and HEV could lead to design of antiviral drugs against both viruses. The third specific aim of the proposed research is to study the three dimensional structure of both the RUB capsid protein by X-ray crystallography and of the RUB virion by cryo-electron microscopy in collaboration with scientists at Purdue University who specialize in structural biology.

风疹病毒（RUB）是主要的人类病原体。 在期间摩擦感染 怀孕会导致严重的先天缺陷（先天性风疹综合征或 CRS）。 风疹在美国通过使用现场，衰减来控制 疫苗。 但是，疫苗接种计划面临两个挑战。 首先，该病毒在世界上的大部分地区都是普遍的，需要 继续保持警惕的疫苗接种努力，以防止重新引入 发生在1989年至1991年之间。其次，在成年女性中，疫苗具有 与慢性关节炎有关 疫苗病毒的持久性。 Rub的另一个重要医疗方面 是CRS患者患糖尿病发病率很高，提供 特定人类病毒与自身免疫性疾病的最佳关联。 拟议研究的长期目标是对 摩擦的分子生物学。 尽管具有医学的重要性，但在 摩擦的分子生物学一直很慢，主要是因为摩擦重复 慢慢地到细胞培养中的低滴度。 最近的地标活动 RUB的分子生物学是“传染性克隆”的发展。 一个 感染性克隆是包含RNA病毒基因组的完整cDNA副本 在质粒中，可以合成感染性RNA转录本在该质粒中 体外。 拟议研究的第一个具体目的是使用 传染性克隆两端：1）。通过定向诱变进行研究 基因组和编码蛋白的特定区域的功能， 和2）。根据现场直播，开发一种传染性克隆 疫苗病毒。 疫苗感染克隆将被引入 “遗传免疫”载体，一种基因组RNA的质粒 在单独用重组质粒转染的细胞中表达， 引发病毒复制。 这个重组向量可能是 可用于全球疫苗接种计划的DNA疫苗 反对摩擦。 疫苗感染克隆也可以用于修饰 避免成年女性遇到的并发症的疫苗。 这 第二个具体目的是研究负责处理的蛋白酶 病毒复制酶或非结构蛋白（NSP）。 作为其中的一部分 努力，乙型肝炎病毒（HEV）的NSP开放阅读框（ORF1）将 表示要分析其处理。 计算机对齐已揭示 摩擦和HEV的NSP孔彼此之间更加紧密相关 而不是其他病毒，尽管摩擦和HEV结构差异差异。 HEV是非A，非B肝炎的肠道传播的主要原因 并且与孕妇的高死亡率有关。 研究 摩擦和HEV的NSP蛋白酶可能导致抗病毒药物的设计 反对这两种病毒。 拟议研究的第三个具体目的是 研究两种摩擦衣壳蛋白的三维结构 X射线晶体学和通过低温电子显微镜在摩擦的摩擦式晶体学中 与专门从事普渡大学的科学家合作 结构生物学。