MATRIX PROTEINS OF ENVELOPED VIRUSES

负链病毒的外周基质（M）蛋白，如 水泡性口炎病毒（VSV）在病毒粒子组装中发挥关键作用， 转录调控 我们建议将分子基础 对于膜结合，核衣壳结合，和 VSV M蛋白的转录抑制功能通过以下方式实现 化学、免疫、遗传和生物物理方法： 1.通过特定化学品和蛋白酶切割产生的肽 将被测序，绘制和测试，沿着合成寡肽， 它们结合磷脂囊泡和核衣壳的能力， 来抑制病毒的转录 2.已经制备了抗M蛋白的单克隆抗体，并将进行检测 因为它们与特定肽的反应性和它们的能力， 抑制M蛋白及其多肽的多种生物学功能。 3.互补组III温度敏感突变M蛋白， 限制在转录抑制，和逆转录M蛋白将是 测试了它们结合脂质囊泡、核衣壳和 特异性单克隆抗体。 4.突变体、回复突变体和野生型M蛋白的重组DNA克隆 将制备基因并对其cDNA进行测序，以确定 与囊泡和核衣壳结合缺陷的M蛋白病变， 转录抑制 我们最终希望制备出 表达载体中的M基因以产生定点缺陷M蛋白。 5.野生型和突变型M蛋白的生物物理学研究及其应用 将进行肽的测定以测定膜动力学的扰动 通过与膜反应的荧光去极化测量， 通过圆二色性光谱测定。 6. VSV-New Jersey和A/WSN/33流感病毒的M蛋白及其生物学特性 单克隆抗体也将通过这些方法进行比较 与M蛋白的功能和进化相关性， 印第安纳州VSV。

The peripheral matrix (M) protein of negative-strand viruses, such as vesicular stomatitis virus (VSV), plays a key role in virion assembly and transcription regulation. We propose to characterize the molecular basis for the membrane binding, nucleocapsid binding, and transcription-inhibition functions of the VSV M protein by the following chemical, immunological, genetic, and biophysical methods: 1. Peptides generated by cleavage with specific chemicals and proteases will be sequenced, mapped, and tested, along with synthetic oligopeptides, for their capacity to bind to phospholipid vesicles and nucleocapsids, and to inhibit viral transcription. 2. Monoclonal antibodies to M protein have been prepared and will be tested for their reactivity with specific peptides and for their capacity to inhibit the various biological functions of M protein and its peptides. 3. Complementation group III temperature-sensitive mutant M proteins, restricted in transcription-inhibition, and revertant M proteins will be tested for their capacity to bind to lipid vesicles, nucleocapsids, and specific monoclonal antibodies. 4. Recombinant DNA clones of mutant, revertant, and wild-type M-protein genes will be prepared and their cDNAs sequenced to identify the sites of lesions in M proteins defective in binding to vesicles and nucleocapsid and in transcription-inhibition. We eventually hope to prepare cDNA clones of M genes in expression vectors to make site-directed defective M proteins. 5. Biophysical studies with wild-type and mutant M proteins and their peptides will be performed to assay perturbations of membrane dynamics measured by fluorescence depolarization reacting with membranes as determined by circular dichroism spectroscopy. 6. M proteins of VSV-New Jersey and A/WSN/33 influenza virus and their monoclonal antibodies will also be examined by these methods for comparison with the functional and evolutionary relatedness of the M protein of VSV-Indiana.