Scheuring U J, Corbeil J, Mosier D E, Theofilopoulos A N
Department of Immunology, Scripps Research Institute, La Jolla, California 92037, USA.
AIDS. 1998 Apr 16;12(6):563-70. doi: 10.1097/00002030-199806000-00004.
Characterization of the effects of infection with HIV-1 on cellular gene expression.
Differential RNA display was applied to compare uninfected and HIV-1LAI-infected CEM cells 24 h post-inoculation. Differential bands were selected, cloned and several clones per band were sequenced. RNase protection assay was used to confirm differential display findings in HIV-1LAI-infected CEM cells as well as in another T-cell line (H9) infected with a different strain (HIV-1 SF33)
Twelve differentially expressed bands, six up- and six downregulated in HIV-infected cells compared with controls, were selected. Four of the six upregulated bands were HIV transcripts. RNase protection assay of the remaining eight bands confirmed differential expression of four genes, including induction of a mariner transposase and moesin as well as suppression of alpha-nascent polypeptide-associated complex and mitochondrial heat shock protein 75 in HIV-1-infected cell cultures. Furthermore, a significant increase of glioma pathogenesis-related protein was found by RNase protection assay.
Based on this initial limited differential display analysis, it was estimated that expression of 3% of the host genes was altered by HIV-1. Amongst the identified gene modifications, the induction of a mariner transposase may alter cellular gene expression itself, whilst the enhanced expression of glioma pathogenesis-related protein suggests a role in the host cell response to viral infection. The increase in moesin may facilitate viral budding and uptake. Furthermore, the suppression of alpha-nascent polypeptide-associated complex may promote translocation of HIV-1 polypeptides into the endoplasmic reticulum, whereas the downregulation of mitochondrial heat shock protein 75 may contribute to a cytopathic effect on mitochondria and possibly impairs antigen presentation.
表征HIV-1感染对细胞基因表达的影响。
应用差异RNA显示技术，比较接种24小时后未感染和感染HIV-1 LAI的CEM细胞。选择差异条带，进行克隆，并对每个条带的多个克隆进行测序。采用核糖核酸酶保护试验，在感染HIV-1 LAI的CEM细胞以及感染不同毒株（HIV-1 SF33）的另一种T细胞系（H9）中，确认差异显示结果。
选择了12条差异表达条带，与对照相比，HIV感染细胞中有6条上调，6条下调。6条上调条带中的4条是HIV转录本。对其余8条带进行核糖核酸酶保护试验，确认了4个基因的差异表达，包括水手转座酶和埃兹蛋白的诱导，以及HIV-1感染细胞培养物中α新生多肽相关复合物和线粒体热休克蛋白75的抑制。此外，通过核糖核酸酶保护试验发现神经胶质瘤发病相关蛋白显著增加。
基于这一初步的有限差异显示分析，估计3%的宿主基因表达因HIV-1而改变。在已确定的基因修饰中，水手转座酶的诱导可能会改变细胞基因表达本身，而神经胶质瘤发病相关蛋白的表达增强表明其在宿主细胞对病毒感染的反应中起作用。埃兹蛋白的增加可能促进病毒出芽和摄取。此外，α新生多肽相关复合物的抑制可能促进HIV-1多肽向内质网的转运，而线粒体热休克蛋白75的下调可能导致对线粒体的细胞病变效应，并可能损害抗原呈递。
