VIRUSES: FROM PARTICLES TO PROTEOMICS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 作为多组分复合物的一部分的溶液相蛋白质运动很少从高分辨率三维结构中明显看出。 蛋白质功能与动力学密切相关，因此了解超分子复合物运动的频率、范围和协调对于理解它们如何发挥作用至关重要。 在这个项目中，病毒是研究超分子复合物中蛋白质动力学的范例。 在过去的一年里，在病毒颗粒的生物物理特性的研究方面取得了重大进展。 目前正在研究的系统包括B型肝炎病毒、腺相关病毒、犬细小病毒、豇豆褪绿斑驳病毒。 最近发表的一篇关于B型肝炎病毒的文章首次测量了兆道尔顿复合物中蛋白质运动的速率和平衡。 在理解与细胞进入相关的pH水平如何调节细小病毒的稳定性和动力学方面也取得了突破性进展。 利用石英晶体微天平和原子力显微镜，测定了不同构象病毒衣壳的粘弹性。 病毒是专性细胞病原体，因此许多细胞蛋白对于病毒感染、复制和从宿主细胞释放至关重要。 利用基于蛋白质组学的方法，Bothner博士正在寻求识别在感染过程中被病毒劫持的细胞蛋白质。 这项工作的意义是双重的，可以阐明病毒的基本生物学，并确定抗病毒剂的新靶点。 Bothner博士关于小鼠RAW细胞的鼠诺如病毒感染的工作导致了线粒体上游半胱氨酸蛋白酶的鉴定，其参与感染。 令人惊讶的是，在感染期间一般抑制半胱天冬酶导致快速的非凋亡性死亡。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The solution-phase protein motion that is part of a multi-component complex is rarely obvious from the high resolution three-dimensional structure. Protein function is intimately connected to dynamics and therefore knowledge of the frequency, range, and coordination of motion by supramolecular complexes is critical to understanding how they function. In this project, viruses are a paradigm for studying protein dynamics in supramolecular complexes. In the past year, significant steps have been made in the studies on the biophysical properties of virus particles. Systems currently under investigation include Hepatitis B virus, Adeno-associated virus, Canine Parvovirus, Cowpea Chlorotic Mottle Virus. A recent publication on Hepatitis B virus presents the first measurements of the rates and equilibria for protein motion in a megadalton complex. Breakthroughs have also been made in understanding how pH levels relevant to cell entry modulate the stability and dynamics of parvoviruses. Through the use of a quartz crystal microbalance and atomic force microscopy, the viscoelastic properties of a virus capsid in different conformation have been determined. Viruses are obligate cellular pathogens and therefore many cellular proteins are critical for viral infection, replication, and release from a host cell. Using proteomics-based approaches, Dr. Bothner is seeking to identify cellular proteins that are hijacked by viruses during the infection process. The significance of this work is two fold; the basic biology of viruses can be elucidated and novel targets for antiviral agents will be identified. Dr. Bothner's work on murine norovirus infection of mouse RAW cells has led to the identification of cysteine proteases upstream of the mitochorndria, which are involved in infection. Surprisingly, inhibition of caspases in general during infection leads to rapid, non-apoptotic death.