THE ROLE OF INTRACELLULAR TRAFFIC IN HIV INFECTION

人类推定的T细胞受体CD_4的研究 免疫缺陷病毒，已经表明这种表面表达 蛋白I被衣霉素阻断，衣霉素是一种有效的蛋白抑制剂 糖基化，在表面受体交替的条件下 都不会受到影响。急性淋巴母细胞瘤的初步研究 通过成功的转基因，白血病细胞得到了延长。 含CD4基因的真核表达载体导入中国仓鼠卵巢 细胞。随后与包含以下内容的另一质粒共转染 多重耐药基因，使分离出 稳定表达大量CD4的克隆。初步数据 在这个材料上表明在CD4中存在复杂或多- 触角杂交寡糖。 已经确定了一个足以 允许0.5x10大小的蛋白质分子通过 核包膜。这一点已经在我们的实验室进行了评估 具有核定位的化学偶联多肽 一种高荧光的细胞质蛋白的序列。该序列 Pro-Lys-Arg-Lys-Va-Cys足以靶向这些 与原子核发生共轭。HIV的TAT和ART基因产物 具有与这个靶向域非常相似的序列。该方法 我们采取的是化学合成相应的多肽 TAT序列检测了该多肽的赋予能力 核本地化。除了这些研究，我们已经开始 检查核孔复合体的结构，穿过它 许多核质交换过程必须发生。这个 利用重组DNA研究了孔道复合体的结构 技巧。主要的核孔糖蛋白np62是 克隆了一大段，并对其进行了测序。

An investigation of CD4, the presumed T-cell receptor for the human immunodeficiency virus, has shown that surface expression of this protei is blocked by tunicamycin, a potent inhibitor of glycosylation, under conditions where alternate surface receptors are unaffected. Initial studies employing acute lymphoblastic leukemic cells have beer extended by the successful transfection of a plasmid containing the cDNA For CD4 into Chinese hamster ovary cells. Subsequent cotransfection with another plasmid containing the multiple drug resistant gene, has permitted the isolation of stable clones expressing large amounts of CD4. Preliminary data on this material indicates the presence in CD4 of complex or multi- antennary hybrid oligosaccharides. A targeting sequence has been identified which is sufficient to allow protein molecules as large as 0.5 x 10 to pass through the nuclear envelope. This has been assessed in our laboratory by chemically coupling peptides having the nuclear localization sequence to a highly fluorescent cytoplasmic protein. The sequence Pro-Lys-Lys-Lys-Arg-Lys-Va-Cys is sufficient to target these conjugates to the nucleus. The tat and art gene products of HIV have sequences very similar to this targeting domain. The approach we have taken is to chemically synthesize peptides corresponding to the tat sequence examined the ability of this peptide to confer nuclear localization. In addition to these studies, we have begun to examine the structure of the nuclear pore complex, across which the many nucleocytoplasmic exchange processes must occur. The structure of the pore complex is explored using recombinant DNA techniques. The major nuclear pore glycoprotein np62 has been cloned and a large segment sequenced.