ROLE OF INTRACELLULAR TRAFFIC IN HIV INFECTION

细胞内运输在 HIV 感染中的作用

• 批准号: 3875584
• 负责人: J A HANOVER
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3875584
• 关键词: AIDS; CD4 molecule; HIV infections; T lymphocyte; acute leukemia; cell nucleus; complementary DNA; cytoplasm; gene expression; glycoproteins; glycosylation; hamsters; human immunodeficiency virus; intracellular membranes; lymphoblast; lymphocytic leukemia; membrane permeability; molecular cloning; neoplastic cell; nucleic acid sequence; ovary; plasmids; pore forming protein; protein sequence; receptor; surface antigens; tissue /cell culture; transfection; tunicamycin; virus infection mechanism; virus receptors; virus replication

CD4, the T-cell receptor for the human immunodeficiency virus, depends upon glycosylation for proper surface expression. Initial studies employing acute lymphoblastic leukemic cells were extended by the successful transfection of a plasmid containing the CDNA for CD4 into mutant and wild type Chinese hamster ovary cells. Characterization of these clones suggests that CD4 contains biantennary unsialylated complex-type oligosaccharides. The data also suggest that the inability of CD4 to reach the cell surface when it lacks carbohydrate is due to the selective degradation of unglycosylated CD4. The REV protein of HIV has been recently shown to lead to the premature transport of the partially spliced MRNA encoding the envelope glycoprotein from the nucleus to the cytoplasm. To develop a.system to examine the effects of REV, a means has been devised for generating nuclei in vitro around exogously added DNA. This method involves using extracts from Xenopus laevis where it has been demonstrated that nuclear assembly can be reconstituted. The availability of such preparations should allow examination of the mechanism of REV function. In other studies, examination has begun of the structure of the nuclear pore complex, across which REV, TAT, and other viral proteins cross following viral infection. The structure of the pore complex is explored using recombinant DNA techniques. The major nuclear pore glycoprotein np62 has been cloned, sequenced and expressed in cultured cells. The gene encoding this protein has been isolated and found to be 2.95 kb in length and devoid of introns. The availability of the primary sequence of this protein has allowed the preparation of anti-peptide antibodies which react with p62. These antibodies should prove useful for probing the function of the pore complex in the HIV life cycle.

人类免疫缺陷病毒的T细胞受体CD4依赖于 糖基化以进行适当的表面表达。初步研究采用 急性淋巴细胞白血病细胞的扩展，成功的 将含有CD4的cDNA的质粒转染到突变体和野生型中， 型中国仓鼠卵巢细胞。这些克隆的特征表明 CD4含有双触角非唾液酸化复合型寡糖。 数据还表明，CD4无法到达细胞表面， 当它缺乏碳水化合物是由于选择性降解， 未糖基化的CD4。 HIV的REV蛋白最近被证明会导致早产儿 编码包膜糖蛋白的部分剪接mRNA的转运 从细胞核到细胞质。开发一个系统来检查 REV的作用，已经设计了一种在体外产生细胞核的方法， 围绕着外来的DNA该方法涉及使用提取物， 非洲爪蟾，已经证明，核组装可以 重组。这种制剂的供应应使 检查REV功能的机制。在其他研究中， 已经开始了核孔复合体的结构，REV， 达特和其他病毒蛋白在病毒感染后交叉。的 使用重组DNA技术探索孔复合物的结构。 主要核孔糖蛋白np62已被克隆，测序， 在培养的细胞中表达。编码这种蛋白质的基因已经被 分离并发现其长度为2.95 kb且缺乏内含子。的 这种蛋白质一级序列的可用性使得 与p62反应的抗肽抗体的制备。这些 抗体应该被证明可用于探测孔复合物的功能 在艾滋病毒的生命周期中。