Chandra P, Vogel A, Gerber T
Cancer Res. 1985 Sep;45(9 Suppl):4677s-4684s.
Compared to other T-lymphotropic human retroviruses, human T-cell leukemia (lymphotropic) virus I (HTLV-I) and HTLV-II, the acquired immunodeficiency syndrome (AIDS)-associated virus, HTLV-III, is a nontransforming cytopathic virus without immortalizing activity. Thus the virus replication is an important event in the manifestation of this disease, and the interruption of viral replication offers an important strategy for the control of AIDS. For this reason we have purified the reverse transcriptase (RT) from HTLV-III and from HTLV-III infected cells to study the structure-activity relationship of RT inhibitors developed in our laboratory. The cellular DNA polymerases from H9 cells were also purified to study the selectivity of RT inhibitors. Purified HTLV-III RT has several distinguishing features: (a) unlike the HTLV-I enzyme it is highly stable and can be kept for several weeks without any loss of activity; (b) using identical procedures of isolation the HTLV-III enzyme shows a much higher activity than does the enzyme from HTLV-I; (c) the Vmax for HTLV-III RT is by severalfold higher than that for the HTLV-I enzyme in the presence of (rC)n X (dG)12 and (rCm)n X (dG)12, and besides the usual template-primers used for RT assay this enzyme has a relatively high affinity for (rAm)n X (dT)12; and (d) the cationic requirements for the transcription of various template-primers are unusual. The purified enzyme has a molecular weight of 95,000-98,000, as judged by the gel filtration method. The purified HTLV-III RT was inhibited by a partially thiolated polycytidylic acid (5-mercaptopolycytidylic acid); the cellular DNA polymerase beta from H9 cells was not sensitive to 5-mercaptopolycytidylic acid. Germanin (synonym, suramin), an antiprotozoan drug, also inhibits HTLV-III RT activity, but the DNA polymerase alpha activity was also sensitive to Germanin. The nonspecific effect of Germanin is probably due to the high content of sulfonic acid residues. This paper describes new approaches for designing specific inhibitors of retroviral reverse transcriptases which may be useful in developing a potential drug against AIDS.
与其他嗜人T淋巴细胞逆转录病毒，即人类T细胞白血病（嗜淋巴细胞）病毒I（HTLV-I）和HTLV-II相比，与获得性免疫缺陷综合征（AIDS）相关的病毒HTLV-III是一种无转化活性的致细胞病变病毒。因此，病毒复制是该疾病表现中的一个重要事件，而中断病毒复制为控制艾滋病提供了一种重要策略。出于这个原因，我们从HTLV-III及其感染的细胞中纯化了逆转录酶（RT），以研究我们实验室开发的RT抑制剂的构效关系。还纯化了H9细胞中的细胞DNA聚合酶，以研究RT抑制剂的选择性。纯化的HTLV-III RT具有几个显著特征：（a）与HTLV-I酶不同，它非常稳定，可以保存数周而活性无任何损失；（b）使用相同的分离程序，HTLV-III酶比HTLV-I酶显示出更高的活性；（c）在存在（rC）nX（dG）12和（rCm）nX（dG）12的情况下，HTLV-III RT的Vmax比HTLV-I酶高几倍，并且除了用于RT测定的常用模板引物外，该酶对（rAm）nX（dT）12具有相对较高的亲和力；（d）各种模板引物转录的阳离子需求不同寻常。通过凝胶过滤法判断，纯化的酶分子量为95,000 - 98,000。纯化的HTLV-III RT被部分硫醇化的聚胞苷酸（5-巯基聚胞苷酸）抑制；H9细胞中的细胞DNA聚合酶β对5-巯基聚胞苷酸不敏感。抗寄生虫药物德国宁（同义词，苏拉明）也抑制HTLV-III RT活性，但DNA聚合酶α活性对德国宁也敏感。德国宁的非特异性作用可能是由于其磺酸残基含量高。本文描述了设计逆转录病毒逆转录酶特异性抑制剂的新方法，这可能有助于开发一种潜在的抗艾滋病药物。
