MOLECULAR DETERMINANTS OF ALPHAVIRUS INDUCED APOPTOSIS

描述：病毒诱导的细胞凋亡对许多人来说很可能是重要的 病毒在其发病机制中的作用，但病毒和细胞因素介导 人们对细胞凋亡知之甚少。贝丝·莱文博士的长期目标 S的研究是为了了解包膜病毒诱导 细胞凋亡。在这里，她建议使用辛德比斯病毒(SIND)，这是原型 甲型病毒，研究病毒诱导的几种哺乳动物细胞凋亡 细胞系和小鼠脑内，以阐明其分子机制。 并探讨细胞凋亡在SIN诱导的脑炎中的作用。 她的初步数据表明，SIN E2包膜糖蛋白 诱导细胞凋亡，SIN诱导的细胞凋亡可以被一种基因阻断 细胞ICE样半胱氨酸蛋白酶活性的抑制物。她的特别之处 目的是确定介导SIN诱导的病毒和细胞因素 细胞凋亡。她将测试SIN E2跨膜蛋白的假设 结构域(TMD)作用于内质网，通过以下途径启动细胞凋亡：1) 识别SIN E2中的死亡结构域，以及2)定义SIN的角色 野生型病毒感染过程中细胞死亡的E2TMD 致死性脑炎的发病机制。她将检验这一假设 细胞ICE样半胱氨酸蛋白酶通过以下途径介导SIN诱导的细胞凋亡：1) 使用半胱氨酸蛋白酶抑制剂，2)监测ICE样活性 感染细胞中的半胱氨酸蛋白酶，以及3)监测相互作用 ICE样半胱氨酸蛋白酶与抗细胞凋亡产物之间的关系 基因、bc1-xl。最后，她将使用酵母双杂交系统来鉴定 大脑中与抗凋亡基因相互作用的基因产物 产品CRMA。

DESCRIPTION: Virally-induced apoptosis is most likely important for many viruses in their pathogenesis, however viral and cellular factors mediating apoptosis are poorly understood. The long-term objective of Dr. Beth Levine s research is to understand the mechanisms by which enveloped viruses induce apoptosis. Here, she proposes to use Sindbis virus (SIN), the prototype alphavirus, to investigate virally-induced apoptosis in several mammalian cell lines and in mouse brains, to elucidate the molecular mechanism of apoptosis and to address the role of apoptosis in SIN-induced encephalitis. Her preliminary data demonstrates that the SIN E2 envelope glycoprotein induces apoptosis and that SIN-induced apoptosis can be blocked by a genetic inhibitor of cellular ICE-like cysteine protease activity. Her specific aims are to identify viral and cellular factors that mediate SIN-induced apoptosis. She will test the hypothesis that the SIN E2 transmembrane domain (TMD) acts in the endoplasmic reticulum to initiate apoptosis by: 1) identifying the death domain in SIN E2, and 2) defining the role of the SIN E2 TMD in cell death during wild-type virus infection and in the pathogenesis of fatal encephalitis. She will test the hypothesis that cellular ICE-like cysteine proteases mediate SIN-induced apoptosis by: 1) using cysteine protease inhibitors, 2) monitoring the activity of ICE-like cysteine proteases in infected cells, and 3) monitoring the interaction between ICE-like cysteine proteases and the product of the anti-apoptotic gene, bcl-xL. Finally, she will use the yeast two hybrid system to identify gene products in the brain that interact with the anti-apoptotic gene product crmA.