The kappa B sites in the human immunodeficiency virus type 1 long terminal repeat enhance virus replication yet are not absolutely required for viral growth
The kappa B sites in the human immunodeficiency virus type 1 long terminal repeat enhance virus replication yet are not absolutely required for viral growth
复制标题
DOI：
10.1128/jvi.71.7.5495-5504.1997
复制
发表时间：
1997-07-01
影响因子：
5.4
通讯作者：
Baltimore, D
中科院分区：
文献类型：
--
作者：
Chen, BK;Feinberg, MB;Baltimore, D
HIV-1对NF-κ B B结合位点的依赖性通过用含有κ B位点增强子突变的HIV-1报告病毒感染细胞，病毒转录用表达内切酶的HIV-1通过流式细胞术分析HIV-1进行单轮感染来测量，1表达胎盘碱性磷酸酶（FLAP）或绿色荧光蛋白（GFP）。表达PLAP和GFP的病毒都在细胞间传播，并允许分析单细胞中的病毒基因表达模式。用不同基础水平的NF-κ B感染一组T细胞系，证明了组成型核NF-κ B的量与野生型病毒优于κ B位点突变体的程度之间的直接相关性，一个具有组成性高水平核NF-κ B的T细胞系，PM 1，当其κ B位点突变时，显示转录降低20倍。相反，在具有低基础水平NF-κ B的T细胞系，SupT 1，增强子中κ B位点的突变对病毒转录或生长速率没有影响，植物血凝素激活的外周血单核细胞显示出对κ B位点的极大依赖性以获得最佳病毒生长。没有标记基因的病毒证实了在具有高基础水平NF-κ B的细胞中κ B位点的突变损害病毒生产的发现。这些数据表明在T细胞中，HIV-1可以利用NF-κ B来增强其生长，但是病毒显然能够在其缺失的情况下生长。
The dependence of human immunodeficiency virus type 1 (HIV-1) on its NF-kappa B binding sites (kappa B sites) for replication in transformed and primary T-cell targets was examined by infecting cells with HIV-1 reporter viruses containing kappa B Site enhancer mutations, Viral transcription was measured either with luciferase-expressing HIV-1 that infects for a single round off by flow cytometric analyses with HIV-1 expressing placental alkaline phosphatase (FLAP) or green-fluorescent]protein (GFP). Both PLAP- and GFP-expressing viruses spread from cell to cell and allowed analysis of viral gene expression patterns in single cells, Infection of a panel of T-cell lines with different basal levels of NF-kappa B demonstrated a direct correlation between the amount of constitutive nuclear NF-kappa B and the degree to which a wild-type virus outperformed kappa B Site mutants, One T-cell line with a constitutively high level of nuclear NF-kappa B, PM1, showed a 20-fold decrease in transcription when its kappa B sites were mutated, In contrast, in a T-cell line with a low basal level of NF-kappa B, SupT1, mutation of the kappa B Site in the enhancer had no effect on viral transcription or growth rate, Phytohemagglutinin-activated peripheral blood mononuclear cells showed a large dependence on the kappa B sites for optimal virus growth. Viruses without marker genes corroborated the finding that mutations to the kappa B sites impair virus production in cells with a high basal level of NF-kappa B, These data show that in T cells, HIV-1 can use NF-kappa B to enhance its growth but the virus is clearly able to grow in its absence.