INCORPORATION OF 12-METHOXYDODECANOATE INTO THE HUMAN IMMUNODEFICIENCY VIRUS-1 GAG POLYPROTEIN PRECURSOR INHIBITS ITS PROTEOLYTIC PROCESSING AND VIRUS PRODUCTION IN A CHRONICALLY INFECTED HUMAN LYMPHOID-CELL LINE
INCORPORATION OF 12-METHOXYDODECANOATE INTO THE HUMAN IMMUNODEFICIENCY VIRUS-1 GAG POLYPROTEIN PRECURSOR INHIBITS ITS PROTEOLYTIC PROCESSING AND VIRUS PRODUCTION IN A CHRONICALLY INFECTED HUMAN LYMPHOID-CELL LINE
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DOI：
10.1073/pnas.88.6.2055
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发表时间：
1991-03-01
影响因子：
11.1
通讯作者：
GORDON, JI
中科院分区：
文献类型：
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作者：
BRYANT, ML;RATNER, L;GORDON, JI
肉豆蔻酸（十四酸；14:0）与人类免疫缺陷病毒 1 (HIV-1) 55-kDa gag 多蛋白前体 (Pr55gag) 的 NH2 末端甘氨酸残基的共价连接对于其蛋白水解加工和病毒组装是必需的。 我们最近发现，亚甲基被单个氧或硫原子取代的肉豆蔻酸酯的几种类似物是酿酒酵母和哺乳动物肉豆蔻酰辅酶A：蛋白N-肉豆蔻酰转移酶（EC 2.3.1.97；NMT）的底物，尽管它们的疏水性降低。 有些药物可抑制急性感染的 CD4+ H9 细胞中的 HIV-1 复制，而不伴随细胞毒性。 为了检查其抗病毒作用的机制，我们用两种类似物 12-甲氧基十二烷酸酯（13-oxamyristate；13-OxaMyr）和 5-辛氧基戊酸酯（6-oxamyristate；6-OxaMyr）进行了标记研究，前者在阻止病毒产生方面比后者更有效。 当 Pr55gag 与酿酒酵母 NMT 在大肠杆菌中共表达时，[H-3]Myristate 和 [H-3]13-OxaMyr 被整合到 Pr55gag 中，效率相当。 [H-3]6-OxaMyr 未掺入，尽管其体外底物性质与 13-OxaMyr 和肉豆蔻酸酯类似。 在慢性感染的 H9 细胞中，[H-3]13-OxaMyr（而非 [H-3]6-OxaMyr）也被有效整合到 HIV-1 Pr55gag 和 nef（阴性因子）中。 类似物掺入使 Pr55gag 从膜重新分布到胞浆部分，并显着减少病毒蛋白酶对其的蛋白水解加工。 13-OxaMyr 和 3'-azido-3'-deoxythymidine (AZT) 协同作用，减少急性感染的 H9 细胞中的病毒产生。 与 AZT 不同，该类似物能够抑制慢性感染的 H9 细胞中的病毒产生（高达 70%）。 而且抑制效果可持续6-8天。 这些结果表明 (i) 其作用机制与 AZT 不同，并且涉及病毒组装的后期步骤； (ii) 类似物可以减少影响病毒复制所需的 AZT 剂量； (iii)类似物和HIV-1蛋白酶抑制剂的组合可能对Pr55gag的加工具有协同作用。
Covalent linkage of myristate (tetradecanoate; 14:0) to the NH2-terminal glycine residue of the human immunodeficiency virus 1 (HIV-1) 55-kDa gag polyprotein precursor (Pr55gag) is necessary for its proteolytic processing and viral assembly. We have shown recently that several analogs of myristate in which a methylene group is replaced by a single oxygen or sulfur atom are substrates for Saccharomyces cerevisiae and mammalian myristoyl-CoA:protein N-myristoyltransferase (EC 2.3.1.97; NMT) despite their reduced hydrophobicity. Some inhibit HIV-1 replication in acutely infected CD4+ H9 cells without accompanying cellular toxicity. To examine the mechanism of their antiviral effects, we performed labeling studies with two analogs, 12-methoxydodecanoate (13-oxamyristate; 13-OxaMyr) and 5-octyloxypentanoate (6-oxamyristate; 6-OxaMyr), the former being much more effective than the latter in blocking virus production. [H-3]Myristate and [H-3]13-OxaMyr were incorporated into Pr55gag with comparable efficiency when it was coexpressed with S. cerevisiae NMT in Escherichia coli. [H-3]6-OxaMyr was not incorporated, even though its substrate properties in vitro were similar to those of 13-OxaMyr and myristate. [H-3]13-OxaMyr, but not [H-3]6-OxaMyr, was also efficiently incorporated into HIV-1 Pr55gag and nef (negative factor) in chronically infected H9 cells. Analog incorporation produced a redistribution of Pr55gag from membrane to cytosolic fractions and markedly decreased its proteolytic processing by viral protease. 13-OxaMyr and 3'-azido-3'-deoxythymidine (AZT) act synergistically to reduce virus production in acutely infected H9 cells. Unlike AZT, the analog is able to inhibit virus production (up to 70%) in chronically infected H9 cells. Moreover, the inhibitory effect lasts 6-8 days. These results suggest that (i) its mechanism of action is distinct from that of AZT and involves a late step in virus assembly; (ii) the analog may allow reduction in the dose of AZT required to affect viral replication; and (iii) combinations of analog and HIV-1 protease inhibitors may have synergistic effects on the processing of Pr55gag.