ISOLATION AND ANALYSIS OF MURINE LEUKEMIA VIRUS RECEPTOR

确定了小鼠C型逆转录病毒的生态宿主范围 通过一种特定的病毒受体的表达。这样做的目的是 建议了解病毒受体的分子基础 在感染中起作用。具体目标是： 1.完成全长Eotroit病毒受体的克隆 吉恩。一个完整基因在不允许的人眼内的表达 细胞将提供生态寄主范围。 2.确定受体表达是否是唯一的决定因素 通过将受体基因导入广谱的 真核物种的范围和感染测试。 3.探讨受体结构与结构之间的关系 和功能。 A.确定转染的小鼠基因是否编码 直接或通过指定修改 人体内源性蛋白质。 B.将编码区的核苷酸序列与 序列库来确定受体基因是否已经 鉴定为一种细胞蛋白质。 C.建立与膜相关的受体结构模型。 识别可能参与包膜的胞外结构域(S) 有约束力的。 D.通过制备抗合成物的抗血清来测试这个模型 来自预测的胞外区的多肽。 1.测试这些抗血清阻断包膜结合和 感染。 2.演示病毒包膜蛋白之间的直接相互作用 (Gp70)和受体通过免疫共沉淀。 3.研究膜gp70-受体相互作用在细胞周期调控中的作用 干扰。 E.确定病毒包膜-受体相互作用的特定位置。 该方法将在绑定中创建空突变 然后选择突变的病毒， 互补性。 这些研究将有助于阐明基因的分子决定因素。 逆转录病毒宿主范围、逆转录病毒进入的机制 小区，以及干扰现象。此外，他们还将 识别逆转录病毒利用的一种细胞蛋白 这是它们生命周期中必不可少的一步。在人类逆转录病毒中 相互作用已被证明在 艾滋病和逆转录病毒诱导的癌症的发病机制。

Ecotropic host range in murine type C retroviruses is determined by the expression of a specific virus receptor. The goal of this proposal is to understand the molecular basis for viral receptor function in infection. The specific aims are to: 1. Complete the cloning of an intact ecotropit virus receptor gene. Expression of an intact gene in non-permissive human EJ cells will confer ecotropic host range. 2. Determine if receptor expression is the sole determinant of ecotropic host range by introducing the receptor gene into a broad range of eucaryotic species and testing for infection. 3. Investigate the relationship between the receptor structure and function. a. Determine if the transfected murine gene encodes the receptor directly or by specifying the modification of an endogenous human protein. b. Compare the nucleotide sequence of the coding region to a sequence bank to determine if the receptor gene has been identified as a cellular protein. c. Model receptor structure with respect to the membrane. Identify extracellular domain(s) that may be involved in envelope binding. d. Test this model by preparing antisera against synthetic peptides from the predicted extracellular domains. 1. Test the ability of these antisera to block envelope binding and infection. 2. Demonstrate direct interaction between virus envelope protein (gp70) and receptor by co-immunoprecipitation. 3. Investigate the role of membrane gp70-receptor interaction in interference. e. Identify specific sites of virus envelope-receptor interaction. The approach will be to create null mutations in the binding domain of the receptor and then to select mutant viruses that complement. These studies will help elucidate the molecular determinants of retrovirus host range, the mechanism of retrovirus entry into cells, and the phenomenon of interference. Further, they will identify a cellular protein that is utilized by retroviruses for an essential step in their lifecycle. In human retroviruses these interactions have been shown to play an important role in the pathogenesis of AIDS and retrovirus-induced cancer.