HUMAN CYTOMEGALOVIRUS INTERACTIONS WITH CELLULAR P53

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，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类巨细胞病毒（HCMV）的婴儿发病率和死亡率发展背后的机制。在过去的7年里，我们已经证明HCMV与关键的细胞周期调节蛋白p53相互作用，并在病毒复制中心隔离它，以及几个关键的DNA损伤蛋白。我们最近表明，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p53的一致结合位点。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蛋白与输入病毒DNA的早期共定位。结合我们在p53敲除细胞中所做的观察，发现病毒滴度急剧下降，病毒DNA复制的延迟和减少以及病毒蛋白表达的延迟，我们认为p53在感染HCMV后的早期和晚期都起着重要作用。确定这些积极作用是本建议的主要重点。我们提出了实现这一目标的三个具体目标。AIM 1是对HCMV感染后p53即刻早期激活的一项研究。AIM 2将决定p53在感染后早期病毒循环中的作用。AIM 3是对p53作为病毒基因表达转录激活因子作用的测定。我们相信，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在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.