HIV-1 Vpr displays natural protein-transducing properties: implications for viral pathogenesis.
HIV-1 Vpr displays natural protein-transducing properties: implications for viral pathogenesis.
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DOI：
10.1006/viro.2002.1576
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发表时间：
2002-10
期刊：
影响因子：
3.7
通讯作者：
M. Sherman;U. Schubert;Samuel A. Williams;C. D. de Noronha;J. Kreisberg;P. Henklein;W. Greene
中科院分区：
文献类型：
--
作者：
M. Sherman;U. Schubert;Samuel A. Williams;C. D. de Noronha;J. Kreisberg;P. Henklein;W. Greene
人类免疫缺陷病毒1型(HIV-1)的14 kDa VPR蛋白在逆转录病毒的生命周期中具有多种功能，包括增强未分裂巨噬细胞的病毒复制，诱导增殖的T淋巴细胞G2期停滞，以及调节HIV-1诱导的细胞凋亡。已在HIV感染患者的血清和脑脊液中检测到胞外VPR。然而，目前还不知道这种形式的VPR是否具有生物活性。VPR包含一个羧基末端的富含氨基酸的片段，它与通过一种称为蛋白质转导的过程介导细胞对多肽的能量和受体无关的摄取的结构域是同源的。在HIV-1Tat、单纯疱疹病毒1DNA结合蛋白VP22和果蝇触角同源转录因子中也存在类似的蛋白转导结构域。我们现在展示了生物活性的合成VPR(SVpr)以及VPR-β-半乳糖苷酶融合蛋白的有效转导。然而，与其他转导蛋白相比，VPR的转导不受蛋白质变性的增强，仅靠VPR的羧基末端碱性结构域不足以跨生物膜进行转导。相反，全长的VPR蛋白有效地转导了大量的细胞，导致剂量依赖的G2细胞周期停滞和凋亡。将VPR添加到细胞外培养液中也可以挽救VPR缺陷的HIV-1毒株在人巨噬细胞培养中的复制。因此，天然VPR可能会针对蛋白质转导进行优化，这一特征可能会增强和扩大艾滋病毒感染的病理影响。
The 14-kDa Vpr protein of human immunodeficiency virus type 1 (HIV-1) serves multiple functions in the retroviral life cycle, including the enhancement of viral replication in nondividing macrophages, the induction of G2 cell-cycle arrest in proliferating T lymphocytes, and the modulation of HIV-1-induced apoptosis. Extracellular Vpr has been detected in the sera and cerebral spinal fluid of HIV-infected patients. However, it is not known whether such forms of Vpr are biologically active. Vpr contains a carboxy-terminal basic amino acid rich segment stretch that is homologous to domains that mediate the energy- and receptor-independent cellular uptake of polypeptides by a process termed protein transduction. Similar functional protein-transducing domains are present in HIV-1 Tat, herpes simplex virus-1 DNA-binding protein VP22, and the Drosophila antennapedia homeotic transcription factor. We now demonstrate effective transduction of biologically active, synthetic Vpr (sVpr) as well as the Vpr-beta-galactosidase fusion protein. However, in contrast to other transducing proteins, Vpr transduction is not enhanced by protein denaturation, and Vpr's carboxy-terminal basic domain alone is not sufficient for its transduction across biological membranes. In contrast, the full-length Vpr protein effectively transduces a broad array of cells, leading to dose-dependent G2 cell-cycle arrest and apoptosis. Addition of Vpr into the extracellular medium also rescues the replication of Vpr-deficient strains of HIV-1 in human macrophage cultures. Native Vpr may thus be optimized for protein transduction, a feature that might enhance and extend the pathological effects of HIV infection.