晶体学和功能分析 ESCRT-I / HIV-1 Gag PTAP 相互作用。
Crystallographic and functional analysis of the ESCRT-I /HIV-1 Gag PTAP interaction.
机构信息
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0580, USA.
出版信息
Structure. 2010 Nov 10;18(11):1536-47. doi: 10.1016/j.str.2010.08.010.
Abstract
Budding of HIV-1 requires the binding of the PTAP late domain of the Gag p6 protein to the UEV domain of the TSG101 subunit of ESCRT-I. The normal function of this motif in cells is in receptor downregulation. Here, we report the 1.4-1.6 Å structures of the human TSG101 UEV domain alone and with wild-type and mutant HIV-1 PTAP and Hrs PSAP nonapeptides. The hydroxyl of the Thr or Ser residue in the P(S/T)AP motif hydrogen bonds with the main chain of Asn69. Mutation of the Asn to Pro, blocking the main-chain amide, abrogates PTAP motif binding in vitro and blocks budding of HIV-1 from cells. N69P and other PTAP binding-deficient alleles of TSG101 did not rescue HIV-1 budding. However, the mutant alleles did rescue downregulation of endogenous EGF receptor. This demonstrates that the PSAP motif is not rate determining in EGF receptor downregulation under normal conditions.
摘要
HIV-1 的出芽需要 Gag p6 蛋白的 PTAP 晚期结构域与 ESCRT-I 的 TSG101 亚基的 UEV 结构域结合。该基序在细胞中的正常功能是受体下调。在这里，我们报告了单独的人 TSG101 UEV 结构域以及野生型和突变型 HIV-1 PTAP 和 Hrs PSAP 九肽的 1.4-1.6Å 结构。P(S/T)AP 基序中的 Thr 或 Ser 残基的羟基与 Asn69 的主链形成氢键。将 Asn 突变为 Pro，阻止主链酰胺，会在体外破坏 PTAP 基序结合并阻止 HIV-1 从细胞中出芽。TSG101 的 N69P 和其他 PTAP 结合缺陷等位基因均不能拯救 HIV-1 出芽。然而，该突变等位基因确实挽救了内源性 EGF 受体的下调。这表明 PSAP 基序在正常条件下不是 EGF 受体下调的限速步骤。
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