Lubon H, Ghazal P, Nelson J A, Hennighausen L
Laboratory of Biochemistry and Metabolism, National Institutes of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
AIDS Res Hum Retroviruses. 1988 Oct;4(5):381-91. doi: 10.1089/aid.1988.4.381.
The binding of nuclear proteins and the functional activity of the HIV-LTR enhancer repeats in different cell lines (Jurkat, CEM, H9, U937, Raji, B cells, T47D, HeLa, 293, and HepG2 cells) was investigated in vitro. Five distinct complexes formed with the enhancer repeat have been identified by an electrophoretic mobility shift assay. The distribution of these complexes varied qualitatively and quantitatively between nuclear proteins from different sources. In the extracts tested, transcription of the HIV-LTR 5' deletion mutants (-453/80, -176/80, -117/80, -103/80, -65/80, and -48/80) was initiated correctly. Transcriptional stimulation dependent upon the presence of the enhancer repeat sequences was observed in all nuclear extracts and was highest in Jurkat, Raji, and B cell extracts. The presence of specific factors and the functional activity of the enhancer repeats as well as other regulatory units in a variety of cells indicates limited host-cell restriction of HIV transcription initiation in vitro.
在体外研究了核蛋白的结合以及HIV-LTR增强子重复序列在不同细胞系（Jurkat、CEM、H9、U937、Raji、B细胞、T47D、HeLa、293和HepG2细胞）中的功能活性。通过电泳迁移率变动分析已鉴定出与增强子重复序列形成的五种不同复合物。这些复合物的分布在来自不同来源的核蛋白之间在定性和定量上有所不同。在所测试的提取物中，HIV-LTR 5'缺失突变体（-453/80、-176/80、-117/80、-103/80、-65/80和-48/80）的转录正确起始。在所有核提取物中均观察到依赖于增强子重复序列存在的转录刺激，并且在Jurkat、Raji和B细胞提取物中最高。特定因子的存在以及增强子重复序列以及各种细胞中其他调节单元的功能活性表明体外HIV转录起始的宿主细胞限制有限。
