Structural Studies of Virus and Receptor Interaction

描述（由申请人提供）：项目摘要：从我目前的博士后职位推进工作是更直接的目标，在本申请中描述为两个具体目标。相对于第一个目标，不同的密切相关的病毒已经适应了以各种方式结合相同的宿主蛋白质，这表明趋同进化为病毒提供了优势。病毒结合这种蛋白质的适应性也改变了致病性。使用结构方法绘制病毒表面相互作用区域可以深入了解受体使用的致病后果，并补充现有的突变研究。第二个目的是针对理解宿主蛋白质的不对称使用的细小病毒。在体外只有一种可溶性受体分子的结合是出乎意料和令人兴奋的。可能的是，结合受体引起病毒的构象变化，并形成对继续感染至关重要的不对称特征，可能导致基因组的脱壳。或者，小的二十面体病毒可能不像以前的结构研究所建议的那样完全对称。在结构测定中广泛使用的平均技术可能最终隐藏了这种病毒和其他病毒系统中功能性不对称特征的存在。目的是研究这两种可能性。在我的博士工作中，对病毒组装的研究导致了一个博士后，在那里我学习了cryoEM重建技术和X射线晶体学。本申请中概述的项目将使用这两种技术，并提供深入研究重建程序套件的机会，过去的理论知识以及如何使用特定程序。要在解决问题方面真正具有创造性，需要对特定算法和例程有全面的计算理解，以便根据手头的数据量身定制重建过程，并获得最高质量的结果。这段过渡期将使我能够继续在学术环境中对病毒结构进行独立研究的最终目标，为该领域做出有价值的贡献，并提供机会传递我所灌输的技能和对科学的热情。相关性：小核糖核酸病毒引起严重的心肌炎，无菌性脑膜炎，并与糖尿病有关。本研究将探讨受体结合对致病性的贡献。细小病毒中的受体使用可能导致功能上必要的偏离精确的二十面体对称性。更好地理解这一现象可以为抗病毒药物提供准确的靶点，并在其他系统中具有更广泛的应用。此外，特别是犬细小病毒的研究可能会导致物种跳跃的澄清。

DESCRIPTION (provided by applicant): Project Summary: Advancing the work from my current post doctoral position is the more immediate goal described in this application as two specific aims. Relative to the first aim, different closely related viruses have adapted to bind the same host protein in a variety of ways, suggesting a convergent evolution that offers an advantage to the virus. The viral adaptation to bind this protein also alters pathogenicity. Using a structural approach to map the area of interaction on the surface of the virus may provide insights into the pathogenic consequences of receptor usage, and complement the existing mutational studies. The second aim is directed at understanding the asymmetric use of a host protein by a parvovirus. The binding of only one soluble receptor molecule in vitro was unexpected and exciting. Possibly, binding receptor instigates a conformational change to the virus and formation of an asymmetric feature critical to continue infection, perhaps leading to the uncoating of the genome. Alternatively, the small icosahedral virus may not be as perfectly symmetrical as structural studies have previously suggested. Averaging techniques used widely in structural determinations may have ultimately hidden from view the presence of functional asymmetric features in this virus and other viral systems. The aim is to investigate both possibilities. The study of viral assembly in my doctoral work, led to a post doc where I have learned both cryoEM reconstruction techniques and X-ray crystallography. The projects outlined in this application will use both techniques and provide the opportunity to delve deeper into the program suites for reconstruction, past knowledge of theory and merely how to use particular programs. To be truly creative in problem solving requires a comprehensive computational understanding of the specific algorithms and routines in order to tailor a reconstruction process to the data at hand and achieve the highest quality results. This transition period will allow me to proceed toward the ultimate goal of continuing independent research in viral structure in an academic setting, making worthy contributions to the field, and providing the opportunity to pass on the skills and the enthusiasm for science that have been instilled in me. Relevance: Picornaviruses cause severe myocarditis, aseptic meningitis and are implicated in diabetes. This study will investigate the contribution of receptor binding to pathogenicity. Receptor usage in parvoviruses may lead to functionally necessary deviations from exact icosahedral symmetry. A better understanding of this phenomenon may provide an exact target for antivirals and have broader application in other systems. Furthermore, study of canine parvovirus in particular may lead to clarification of species jumping.