Identification of a human immunodeficiency virus type 1 TAR binding protein in human hepatoblastoma HepG2 cells that trans-activates HIV-1 LTR-directed gene expression.
Identification of a human immunodeficiency virus type 1 TAR binding protein in human hepatoblastoma HepG2 cells that trans-activates HIV-1 LTR-directed gene expression.
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人肝母细胞瘤 HepG2 细胞中人类免疫缺陷病毒 1 型 TAR 结合蛋白的鉴定，该蛋白反式激活 HIV-1 LTR 定向基因表达。
DOI：
10.1089/dna.1994.13.67
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发表时间：
1994
影响因子：
3.1
通讯作者：
Banerjee,R
中科院分区：
文献类型：
--
作者：
Pizzella,T;Banerjee,R
最近，我们发现人类免疫缺陷病毒（HIV-1）长末端重复（LTR）导向的氯霉素乙酰转移酶（CAT）基因在人肝母细胞瘤HepG2细胞中有效表达，这些细胞可以支持HIV-1的高效复制。在这项研究中，我们发现HepG2细胞含有一个与hiv -1反式激活区（TAR）结合的核因子，我们将其命名为HepG2衍生的TAR结合蛋白（HTBP）。使用在TAR区携带不同突变的合成寡核苷酸探针进行凝胶阻滞实验，以及使用这些寡核苷酸进行竞争DNA迁移实验，发现HIV-1 LTR中包含+7至+13个核苷酸（5 ' -TCTGGTT-3 '）的结合位点。使用- 65HIV-1 LTR CAT作为报告质粒和含有这些突变寡核苷酸的各种竞争质粒的Anin vivoCAT竞争实验也表明HTBP可以影响HIV-1ltr定向CAT基因在HepG2细胞中的表达通过与TAR区特定序列的相互作用。
Recently, we have shown that the human immunodeficiency virus (HIV-1) long terminal repeat (LTR) directed chloramphenicol acetyltransferase (CAT) gene is efficiently expressed in human hepatoblastoma HepG2 cells and these cells can support productive HIV-1 replication. In this study we show that HepG2 cells contain a nuclear factor that binds to the HIV-1trans-activating region (TAR), which we named HepG2-derived TAR binding protein (HTBP). Gel retardation assays using synthetic oligonucleotide probes carrying different mutations in the TAR region and competition DNA mobility-shift experiments using these oligonucleotides revealed the binding site encompassing between +7 and +13 nucleotides (5′-TCTGGTT-3′) in the HIV-1 LTR. Anin vivoCAT competition assay using −65HIV-1 LTR CAT as a reporter plasmid and various competitor plasmids containing these mutated oligonucleotides also demonstrated that HTBP can influence the HIV-1 LTR-directed CAT gene expression in HepG2 cells by interaction with a specific sequence in the TAR region.