MOLECULAR DETERMINANTS OF ALPHAVIRUS INDUCED APOPTOSIS

描述：病毒诱导的细胞凋亡对许多 病毒在其发病机制中，然而，病毒和细胞因子介导 细胞凋亡知之甚少。 贝丝·莱文医生的长期目标 的研究是了解包膜病毒诱导的机制 凋亡 在这里，她建议使用辛德比斯病毒（SIN）， α病毒，研究病毒诱导的几种哺乳动物细胞凋亡 细胞系和小鼠脑中，以阐明 细胞凋亡和解决细胞凋亡在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 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抑制剂。 她的具体 目的是确定介导SIN诱导的病毒和细胞因子， 凋亡 她将检验SIN E2跨膜 结构域（TMD）通过以下方式在内质网中起作用以启动细胞凋亡：1） 鉴定SIN E2中的死亡结构域，和2）定义SIN的作用 E2 TMD在野生型病毒感染过程中细胞死亡中的作用以及在 致命性脑炎的发病机制。 她将检验一个假设， 细胞ICE样半胱氨酸蛋白酶通过以下方式介导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样蛋白酶的活性， 感染细胞中的半胱氨酸蛋白酶，和3）监测相互作用 ICE样半胱氨酸蛋白酶和抗凋亡蛋白的产物之间的关系 基因bcl-xL。 最后，她将使用酵母双杂交系统来鉴定 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.