HUMAN CYTOMEGALOVIRUS INTERACTIONS WITH CELLULAR P53

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们工作的长期目标是了解先天性感染人类巨细胞病毒(HCMV)的婴儿发病率和死亡率背后的机制。在过去的7年里，我们已经证明了HCMV与关键的细胞周期调节蛋白P53相互作用，并将其隔离在病毒复制中心内，以及几个关键的DNA损伤蛋白。我们最近发现，这种隔离依赖于P53蛋白上完整的DNA结合域，并在病毒基因组中发现了21个共同的P53结合位点。这些事实使我们假设，P53可能被病毒招募为转录因子。我们还观察到P53蛋白与输入的病毒DNA很早就发生了共存。结合我们在p53基因敲除细胞中所做的观察，发现病毒滴度急剧下降，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在感染HCMV后的早期和晚期都发挥着重要作用。确定这些积极作用是这项建议的主要重点。为了实现这一目标，我们提出了三个具体目标。目的1研究人巨细胞病毒感染对p53基因的即刻早期激活作用。目的2确定P53在感染后早期病毒循环中的作用。目的3确定P53作为转录激活因子对病毒基因表达的作用。我们认为，阐明与关键的细胞周期调节因子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.