Paramyxovirus Entry

描述(由申请人提供)：我们学习的目的是了解 新城疫病毒副粘病毒膜融合的分子机制 以疾病病毒为原型。与大多数副粘病毒系统一样，膜融合 需要HN和F两种蛋白质。我们的研究集中在两个问题上，如何 F蛋白介导膜融合及RN蛋白在膜融合中的作用 进程。我们对F蛋白的突变分析表明，两个七肽 重复结构域，HR1和HR2，在融合活性中起重要作用 蛋白。我们已经证明了具有这些结构域序列的多肽相互作用 但很可能只有在融合激活后才会发生。我们还发现序列 来自HN蛋白的DNA将与F蛋白中的特定结构域相互作用。 此外，我们还在另一个F蛋白七肽中进行了点突变 重复结构域，HR3，它消除了对RN蛋白的绝对需求 核聚变。为了解决这些发现提出的问题，我们提出了四个 具体目的：1)确定HN蛋白和F蛋白相互作用的限度 域名。2)研究多肽与多肽类似物的相互作用 来自HR1、HR2和RN序列，以便阐明参与 F突变体的融合抑制，以确定对 构象位移，并确定RN-F蛋白中重要的残基 互动。3)定义分子中构象转移的性质 糖蛋白利用多肽、多肽类似物以及多肽特异性 抗血清。4)。探讨角色依恋在融合启动中的作用。

DESCRIPTION (provided by applicant): The goal of our studies is to understand the molecular mechanisms of paramyxovirus membrane fusion using Newcastle disease virus as a prototype. As in most paramyxovirus systems, membrane fusion requires both HN and F proteins. Our studies have focused on two questions, how the F protein mediates membrane fusion and the role of RN protein in that process. Our mutational analysis of the F protein has shown that two heptad repeat domains, HR1 and HR2, are important in the fusion activity of the protein. We have shown that peptides with sequences from these domains interact but likely only upon activation of fusion. We have also found that sequences from the HN protein will interact with a specific domain within the F protein. Furthermore, we have made a point mutation within another F protein heptad repeat domain, HR3, which eliminated the absolute requirement for RN protein in fusion. To address the questions raised by these findings we propose four specific aims: 1) To determine limits of HN protein and F protein interacting domains. 2) To characterize the interactions of peptides and peptide analogues from the HR1, HR2, and RN sequences in order to clarify mechanisms involved in fusion inhibition by F mutants, to identify residues important for conformational shifts, and to identify residues important in RN-F protein interactions. 3) To define the nature of conformational shifts in the glycoproteins using peptides, peptide analogues, as well as peptide specific antisera. 4). To explore the role attachment in initiation of fusion.