Cloning and analysis of the promoter region of CCR5, a coreceptor for HIV-1 entry.
Cloning and analysis of the promoter region of CCR5, a coreceptor for HIV-1 entry.
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HIV-1 进入的辅助受体 CCR5 启动子区域的克隆和分析。
DOI：
10.4049/jimmunol.159.11.5441
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发表时间：
1997
影响因子：
4.4
通讯作者：
A. Fauci
中科院分区：
文献类型：
--
作者：
H. Moriuchi;M. Moriuchi;A. Fauci
趋化因子受体 CCR5 是巨噬细胞嗜性 HIV-1 菌株进入细胞的辅助因子。 CCR5 的表达仅限于 T 细胞、巨噬细胞和某些细胞系；然而，控制其表达的机制仍然很大程度上未知。为了描述这些机制，克隆并表征了来自 CCR5 紧邻 5' 上游区域的约 1.0 kb DNA。 CCR5 启动子活性被 PMA 上调，相对于转录起始位点跨越 -417 至 +61 的区域足以满足基础活性和诱导活性。 DNase I 足迹分析显示了该区域内的几个保护区，凝胶迁移分析确定了转录因子 Oct-1、Oct-2、T 细胞因子 1α 和 GATA1 的结合位点。 IL-2 或抗 CD3 Ab 也可诱导 CCR5 启动子活性，而抗 CD28 Ab 刺激可显着降低 CD3 介导的 CCR5 启动子上调。流式细胞术证实了细胞表面表达水平的发现。进一步描述 CCR5 启动子的调控对于更全面地了解 HIV 疾病的发病机制非常重要。
The chemokine receptor CCR5 is a cofactor for cellular entry of macrophage-tropic strains of HIV-1. Expression of CCR5 is restricted to T cells, macrophages, and certain cell lines; however, the mechanisms controlling its expression remain largely unknown. To delineate these mechanisms, approximately 1.0 kb of DNA from the immediate 5' upstream region of CCR5 was cloned and characterized. CCR5 promoter activity was up-regulated by PMA, and a region spanning -417 to +61 relative to the transcription start site was sufficient for the basal and induced activity. DNase I footprinting assays demonstrated several protected areas within this region, and gel shift assays determined binding sites for transcriptional factors Oct-1, Oct-2, T cell factor 1alpha, and GATA1. CCR5 promoter activity was also induced by IL-2 or anti-CD3 Ab, while stimulation with anti-CD28 Ab markedly reduced CD3-mediated up-regulation of the CCR5 promoter. Flow cytometry confirmed the findings at the level of cell surface expression. Further delineation of the regulation of the CCR5 promoter will be important for a more comprehensive understanding of the pathogenesis of HIV disease.