HUMAN CYTOMEGALOVIRUS INTERACTIONS WITH CELLULAR P53

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 我们工作的长期目标是了解先天性感染人巨细胞病毒（HCMV）的婴儿发病率和死亡率发展背后的机制。 在过去的7年中，我们已经表明，HCMV与关键的细胞周期调控蛋白p53相互作用，并将其隔离，以及病毒复制中心内的几个关键DNA损伤蛋白。我们最近表明，这种隔离依赖于p53蛋白上的完整DNA结合结构域，并在病毒基因组中发现了21个p53的共有结合位点。 这些事实使我们假设p53可能被病毒募集为转录因子。我们还观察到p53蛋白与输入病毒DNA的非常早期的共定位。 再加上我们在p53基因敲除细胞中所做的观察，这些观察显示病毒滴度急剧下降，病毒DNA复制延迟和减少以及病毒蛋白表达延迟，我们认为p53在HCMV感染后的早期和晚期都起着重要作用。本建议的主要重点是确定这些积极作用。我们提出三个具体目标来实现这一目标。 目的1是研究HCMV感染后p53的即刻早期激活。AIM 2将确定p53在感染后早期病毒环化中的作用。目的3是确定p53作为病毒基因表达的转录激活因子的作用。我们认为，阐明与关键细胞周期调节因子p53的相互作用可能有助于我们理解先天性感染婴儿中观察到的中枢神经系统表现的发展。

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 long-term goal of our work is to understand the mechanism behind the development of morbidity and mortality in infants congenitally infected with human cytomegalovirus (HCMV). In the last 7 years we have shown that HCMV interacts with the key cell cycle regulatory protein p53 and sequesters it, and several key DNA damage proteins, within the viral replication centers. We have recently shown that this sequestration is dependent upon an intact DNA binding domain on the p53 protein and have found 21 consensus binding sites for p53 within the viral genome. These facts have led us to hypothesize that p53 may be recruited as a transcription factor by the virus. We have also observed very early colocalization of the p53 protein with input viral DNA. Coupled with the observations we have made in p53 knockout cells, which show dramatic decreases in viral titers, delays and decreases in viral DNA replication and delays in viral protein expression, we believe that p53 plays important roles at both early and late times post infection with HCMV. It is the major focus of this proposal to ascertain these active roles. We propose three specific aims to accomplish this goal. AIM 1 is an investigation of the immediate early activation of p53 by HCMV infection. AIM 2 will determine the role of p53 in viral circularization at early times post infection. AIM 3 is a determination of the role of p53 as transcriptional activator for viral gene expression. We believe that elucidating the interactions with the key cell cycle regulator p53 may aid in our understanding of the development of the central nervous system manifestations observed in the congenitally infected infant.