Zolotukhin A S, Harford J B, Felber B K
Human Retrovirus Pathogenesis Group, National Cancer Institute-Frederick Cancer Research and Development Center, MD 21702-1201.
Nucleic Acids Res. 1994 Nov 11;22(22):4725-32. doi: 10.1093/nar/22.22.4725.
Expression of the structural proteins of the human immunodeficiency virus type 1 (HIV-1), the human T-cell leukemia virus type I (HTLV-I), and of the transferrin receptor (TfR) mRNA depends on posttranscriptional regulatory mechanisms involving both positive and negative elements. In these systems the presence of elements decreasing mRNA expression have been demonstrated. The regulatory proteins (Rev, Rex or iron response element binding protein IRE-BP) antagonize the effects of the downregulatory elements by interacting directly with specific mRNA sites (Rev responsive element, RRE, Rex responsive element, RXRE, or iron responsive elements, IREs) resulting in stabilization and efficient expression of the corresponding mRNAs. To investigate whether this strategy involves common pathways of mRNA utilization, we have studied expression from hybrid mRNAs that contained these previously identified HIV-1 or TfR instability determinants and the binding sites of the regulatory proteins Rev, Rex and/or IRE-BP. Our results demonstrate that only low levels of these hybrid mRNAs accumulate in the absence of the positive regulatory factors Rev, Rex or IRE-BP. The presence of these factors counteracts the effect of heterologous downregulatory elements resulting in increased accumulation of the hybrid mRNAs. However, while Rev or Rex regulation also resulted in efficient protein expression, the IRE-BP only affected mRNA levels without significantly affecting protein expression, suggesting that the pathways of mRNA stabilization/expression are different in these systems.
1型人类免疫缺陷病毒(HIV-1)、I型人类T细胞白血病病毒(HTLV-I)的结构蛋白以及转铁蛋白受体(TfR)mRNA的表达取决于涉及正负两种元件的转录后调控机制。在这些系统中,已证实存在降低mRNA表达的元件。调控蛋白(Rev、Rex或铁反应元件结合蛋白IRE-BP)通过直接与特定mRNA位点(Rev反应元件RRE、Rex反应元件RXRE或铁反应元件IRE)相互作用,拮抗下调元件的作用,从而导致相应mRNA的稳定和高效表达。为了研究这种策略是否涉及mRNA利用的共同途径,我们研究了包含这些先前鉴定的HIV-1或TfR不稳定决定因素以及调控蛋白Rev、Rex和/或IRE-BP结合位点的杂交mRNA的表达。我们的结果表明,在没有阳性调控因子Rev、Rex或IRE-BP的情况下,这些杂交mRNA仅积累低水平。这些因子的存在抵消了异源下调元件的作用,导致杂交mRNA积累增加。然而,虽然Rev或Rex调控也导致了有效的蛋白质表达,但IRE-BP仅影响mRNA水平,而没有显著影响蛋白质表达,这表明这些系统中mRNA稳定/表达的途径是不同的。