Dynamics and mechanisms of filovirus envelop glycoproteins

总结 埃博拉疫情日益频繁和严重，需要扩大治疗方案， 预防措施。解决这一未满足的需求将需要更深入地了解分子生物学。 丝状病毒复制的潜在机制。特别是，丝状病毒包膜期间发生的动态事件 糖蛋白（GP）在进入细胞期间介导的膜融合几十年来一直没有得到阐明。 先前的研究已经确定了GP的蛋白水解裂解、受体结合和GP的化学环境。 晚期内体是关键的。但是这些事件和变量促进的分子机制 GP介导的膜融合尚不清楚。因此，丝状病毒融合的完整和特异性模型， 目前还不存在整合宿主因素、环境条件和GP构象变化的方法。 因此，丝状病毒进入仍然是抑制剂未利用的靶标。我们的长期目标是发展一个 GP介导的膜融合的完整机制模型。我们最近的出版物，其中我们 证明了单分子荧光方法在阐明构象动力学的能力， EBOV GP在病毒体表面的表现，证明了我们为此所做的初步努力。我们的目标是在此基础上 通过提出涉及病毒学、细胞学、结构学和生物物理学的多学科研究取得成功 阐明来自多种丝状病毒的GP的动力学和机制的方法。我们将描述 构象变化、宿主因素和环境变量促进丝状病毒的机制 膜融合和进入细胞。完成拟议的研究将提供机械的见解， 丝状病毒进入以及可以用新疗法和免疫原开发的病毒和宿主靶标。

SUMMARY The increasing frequency and severity of Ebola outbreaks demands an expanded repertoire of treatments and preventative measures. Answering this unmet need will require a deeper understanding of the molecular mechanisms underlying filovirus replication. In particular, the dynamic events that occur during filovirus envelope glycoprotein (GP)-mediated membrane fusion during entry into cells have evaded elucidation for decades. Previous studies have identified proteolytic cleavage of GP, receptor binding, and the chemical environment of the late endosome as being critical. But the molecular mechanisms by which these events and variables promote GP-mediated membrane fusion are not known. As a result, a complete and specific model of filovirus fusion, which integrates host factors, environmental conditions, and GP conformational changes currently does not exist. Therefore, filovirus entry continues to be an unutilized target for inhibitors. Our long-term goal is to develop a complete mechanistic model of GP-mediated membrane fusion. Our recent publications, in which we demonstrate the power of single-molecule fluorescence methods in elucidating the conformational dynamics of EBOV GP on the surface of virions, demonstrate our initial efforts toward this end. Here we aim to build on this success by proposing a multidisciplinary study involving virological, cellular, structural, and biophysical methodologies to elucidate the dynamics and mechanisms of GPs from multiple filoviruses. We will characterize the mechanisms by which conformational changes, host factors, and environmental variables facilitate filovirus membrane fusion and entry into cells. Completion of the proposed research will provide mechanistic insights into filovirus entry and the viral and host targets that could be exploited with novel therapies and immunogens.