POLIOVIRUS RECEPTOR AND INTERACTIONS WITH POLIOVIRUS

人脑皮质激素受体相互作用的分子参数 脊髓灰质炎病毒(HPVR)是一种80kD的Ig样细胞表面糖蛋白，具有 一般结构域结构V-C2-C2，以及脊髓灰质炎病毒。 这将涉及到hPVR多肽通过位点定向的特异性变化。 突变和病毒突变体的遗传分析。受体变异体 将在小鼠细胞中表达，用于功能研究。此外, 受体变异体将在CHO细胞中过度表达以产生大量 适合结晶和X射线分析的纯化蛋白的量 分析，并用于受体/病毒的冷冻电子显微镜研究 很复杂。这些结构研究将与M.G.合作。 罗斯曼。脊髓灰质炎病毒感染动物研究中的一个谜团是 受体信使核糖核酸和多肽的广泛表达 与有限的一组目标组织形成对比。以确定基因 负责表达的元件，hPVR基因的启动子将 在组织培养细胞中进行解剖和研究，目的是 分离顺式作用序列和反式作用因子。此外， 受hPVR启动子调控的表达将由生殖系研究 小鼠的转化及不同片段连锁的LacZ基因 HPVR基因启动子。确定hPVR可能的相互作用 通过细胞质结构域与细胞内蛋白质结合，进行遗传筛选 在酵母中使用双杂交系统将被采用。这些研究 将包括hPVR和淋巴细胞归巢之间可能的相互作用 受体CD44。最后，小鼠神经毒性的遗传决定因素 对脊髓灰质炎病毒株PV1(LS-a)的研究将注意 衣壳蛋白编码序列之外的基因组序列。

Molecular parameters of the interaction between the human receptor for poliovirus (hPVR), a 80 kD Ig-like, cell-surface glycoprotein with a general domain structure V-C2-C2, and the poliovirion will e studied. This will involve specific changes of hPVR polypeptide by site-directed mutagenesis, and genetic analyses of viral mutants. Receptor variants will be expressed in mouse cells for functional studies. In addition, receptor variants will be over-expressed in CHO cells to produce large quantities of purified protein suitable for crystallization and X-ray analysis, and for cryoelectron microscopic studies of the receptor/virus complex. These structural studies will be a collaboration with M.G. Rossmann. An enigma in research of poliovirus infection of animals is the apparent widespread expression of receptor mRNA and polypeptides contrasted with a limited set of target tissues. To identify genetic elements responsible for expression, the promoter of the hPVR gene will be dissected and studied in tissue culture cell with the objective to isolate cis-acting sequences and trans-acting factors. Moreover, expression regulated by the hPVR promoter will be studied by germline transformation of mice and the gene for lacZ linked to different segments of the hPVR gene promoter. To determine possible interactions of hPVR with intracellular proteins via the cytoplasmic domains, a genetic screen employing the two-hybrid system in yeast will be employed. These studies will include possible interactions between hPVR and the lymphocyte homing receptor CD44. Finally, the genetic determinants of mouse neurovirulence of PV1(LS-a), a strain of poliovirus, will be studied with attention to genomic sequences outside the coding sequence for the capsid protein.