Lack of a negative influence on viral growth by the nef gene of human immunodeficiency virus type 1.
Lack of a negative influence on viral growth by the nef gene of human immunodeficiency virus type 1.
复制标题
DOI：
10.1073/pnas.86.23.9544
复制
发表时间：
1989-12
期刊：
影响因子：
--
通讯作者：
Sunyoung Kim;K. Ikeuchi;R. Byrn;J. Groopman;D. Baltimore
中科院分区：
文献类型：
--
作者：
Sunyoung Kim;K. Ikeuchi;R. Byrn;J. Groopman;D. Baltimore
摘要 人类免疫缺陷病毒 1 型 (HIV-1) 在其基因组 3' 端包含一个称为 nef 的开放阅读框。据报道，nef 基因产物可通过与长末端重复区域相互作用抑制病毒转录，从而下调病毒生长。我们比较了两种同基因 HIV-1 (HIV-1-WI3) 毒株，其中一种缺乏 nef 表达，发现它们在体外生长方面几乎没有差异。我们通过测量感染后 HIV 特异性低分子量 DNA 和 RNA 来测试对病毒进入、DNA 合成和 RNA 表达的影响。 nef+ 和 nef- 菌株之间 DNA 和 RNA 合成的定性和定量方面具有可比性。还通过感染过程中逆转录酶活性的变化来检查对病毒生长的影响。 nef基因产物的存在未能减缓所测试的几种不同细胞类型中的病毒生长，包括人T淋巴细胞系H9和CEM-SS、富含CD4+细胞的人原代T细胞以及人单核细胞系U-937和THP-1。相反，nef+ 菌株在某些细胞类型中比 nef- 菌株生长更有效。使用不同病毒 HIV-1-432 的 nef+ 和 nef- 毒株也获得了相同的结果，据报道，该病毒的 Nef 对病毒生长有负面影响。我们的数据表明，Nef 蛋白不会充当负面因素，至少在我们研究中使用的实验系统中是这样。
Abstract Human immunodeficiency virus type 1 (HIV-1) contains an open reading frame called nef at the 3' end of its genome. The nef gene product has been reported to down-regulate viral growth by suppressing viral transcription through interaction with the long terminal repeat region. We have compared two isogenic HIV-1 (HIV-1-WI3) strains, one of which lacks nef expression, and found little difference between them in in vitro growth. We tested effects on viral entry, DNA synthesis, and RNA expression by measuring HIV-specific low molecular weight DNA and RNA after infection. The qualitative and quantitative aspects of DNA and RNA synthesis were comparable between the nef+ and nef- strains. The effects on viral growth were also examined by following changes in reverse transcriptase activity during the course of infection. The presence of the nef gene product failed to slow viral growth in several different cell types tested, including the human T-lymphocyte cell lines H9 and CEM-SS, human primary T cells enriched for CD4+ cells, and human monocytic cell lines U-937 and THP-1. On the contrary, the nef+ strain grew more efficiently in some cell types than the nef- strain. The same results were obtained with nef+ and nef- strains of a different virus, HIV-1-432, whose Nef had been reported to have a negative effect on viral growth. Our data suggest that the Nef protein does not act as a negative factor, at least in the experimental systems employed in our studies.