STRUCTURE & EXPRESSION OF THE PARAMYXOVIRUS SV5 GENOME

结构

基本信息

批准号：
3135080
负责人：
ROBERT A LAMB
金额：
$ 21.45万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-01-01 至 1995-12-31
项目状态：
已结题

项目摘要

The structure and function of the genes and proteins of the paramyxovirus SV5 will be investigated with particular emphasis on studying the hemagglutinin-neuraminidase (HN), the fusion (F) and the P and V proteins. The paramyxoviruses are the etiological agents of many biologically and economically important diseases of man and lower animals. The paramyxovirus SV5 is a prototype of the paramyxovirus family of non- segmented negative strand RNA viruses which includes Sendai virus, Newcastle disease virus, mumps virus, human parainfluenza virus types 1-5, the morbillivirus subgroup including measles virus and the pneumovirus subgroup including respiratory syncytial virus. We will determine the site(s) on HN with which the cellular chaperone protein GRP78-BiP interacts and elucidate the mechanism by which SV5-infection of cells induces synthesis of GRP78-BiP. The HN molecule will be engineered to synthesize soluble and secreted HN to understand further the requirement for proper protein folding and oligomerization for intracellular transport. The majority of HN expressed at the cell surface is not incorporated into virions but is internalized and degraded in lysosomes. We will determine the domains and then the precise molecular signals of HN involved in its internalization from the cell surface. We will also elucidate if the HN lysosomal targeting signal is the same as the internalization signal. The rate and extent of the internalization will be determined using biochemical assays and the role of sialic acid as a ligand in the internalization process will be investigated. To understand the cellular pathway by which HN is internalized from the cell surface we will undertake an electron microscopic examination. The oligomerization, intracellular transport, carbohydrate modification and proteolytic processing of the F protein will be investigated. We will examine the role of the conserved residues in the fusion related external domain of the F protein in mediating cell to cell fusion. We will investigate further the paramyxovirus RNA editing process which produces the P and V mRNAs and we will quantitate the frequency of the G nucleotide insertion process at the specific site on the "P" gene. We will determine the subcellular localization of protein V and its relative abundance in virions. An examination will be made of the ability of protein V to coordinate zinc or other heavy metals and if so determine the role of the cysteine residues in this process. An investigation will be made of the ability of P and V to bind to RNA and if so the regions of the proteins involved will be determined.

丙糖病毒的基因和蛋白质的结构和功能 SV5将被调查，特别是研究血凝素 - 神经酶酶（HN），融合（F）和P和V蛋白。帕托病毒是许多生物学和人类和下动物的经济重要疾病。这 Paramyxovirus sv5是非 - 氧病毒家族的原型分段的负链RNA病毒，包括仙台病毒，纽卡斯尔病毒，腮腺炎病毒，人类副帕氏素病毒1-5型病毒包括麻疹病毒和肺炎病毒在内的虫比病毒亚组包括呼吸综合病毒的亚组。我们将确定在HN上与细胞伴侣蛋白GRP78-BIP相互作用的位点并阐明了细胞的SV5感染引起的机制 GRP78-BIP的合成。 HN分子将被设计为合成可溶和分泌的HN，以进一步了解适当的要求蛋白质折叠和寡聚化以进行细胞内转运。这在细胞表面表达的大多数HN未纳入病毒体但被内在化并在溶酶体中降解。我们将确定域，然后涉及HN的精确分子信号从细胞表面进行内在化。我们还将阐明是否HN 溶酶体靶向信号与内部化信号相同。这内部化的速率和程度将使用生化确定测定和唾液酸作为配体在内在化中的作用过程将进行研究。了解细胞途径 HN是从细胞表面内部化的，我们将进行电子显微镜检查。寡聚，细胞内转运， F蛋白的碳水化合物修饰和蛋白水解加工将被调查。我们将研究保守残基在融合相关的F蛋白介导细胞中F蛋白的外部结构域融合。我们将进一步研究Paramyxovirus RNA编辑过程产生P和V mRNA，我们将定量 “ P”基因上特定位点的G核苷酸插入过程。我们将确定蛋白质V及其的亚细胞定位病毒体的相对丰度。考试将对能力进行检查蛋白质v以协调锌或其他重金属，如果确定半胱氨酸残基在此过程中的作用。调查会由P和V与RNA结合的能力制成，如果是这样的区域将确定所涉及的蛋白质。