BACKGROUND Changes in iron metabolism frequently accompany HIV-1 infection. However, while many clinical and in vitro studies report iron overload exacerbates the development of infection, many others have found no correlation. Therefore, the multi-faceted role of iron in HIV-1 infection remains enigmatic. METHODS RT-qPCR targeting the LTR region, gag, Tat and Rev were performed to measure the levels of viral RNAs in response to iron overload. Spike-in SILAC proteomics comparing i) iron-treated, ii) HIV-1-infected and iii) HIV-1-infected/iron treated T lymphocytes was performed to define modifications in the host cell proteome. Data from quantitative proteomics were integrated with the HIV-1 Human Interaction Database for assessing any viral cofactors modulated by iron overload in infected T lymphocytes. RESULTS Here, we demonstrate that the iron overload down-regulates HIV-1 gene expression by decreasing the levels of viral RNAs. In addition, we found that iron overload modulates the expression of many viral cofactors. Among them, the downregulation of the REV cofactor eIF5A may correlate with the iron-induced inhibition of HIV-1 gene expression. Therefore, we demonstrated that eiF5A downregulation by shRNA resulted in a significant decrease of Nef levels, thus hampering HIV-1 replication. CONCLUSIONS Our study indicates that HIV-1 cofactors influenced by iron metabolism represent potential targets for antiretroviral therapy and suggests eIF5A as a selective target for drug development.

中文翻译：

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铁超载下调了感染的T淋巴细胞中HIV-1 Rev辅因子eIF5A的表达。

背景技术铁代谢的变化经常伴随HIV-1感染。但是，尽管许多临床和体外研究报告均表明铁超载加剧了感染的发展，但其他许多研究却没有相关性。因此，铁在HIV-1感染中的多方面作用仍然是个谜。方法进行针对LTR区，gag，Tat和Rev的RT-qPCR来检测铁超负荷时病毒RNA的水平。刺入式SILAC蛋白质组学比较了i）铁处理的，ii）HIV-1感染的和iii）HIV-1感染的/铁处理的T淋巴细胞，以定义宿主细胞蛋白质组中的修饰。来自定量蛋白质组学的数据与HIV-1人类相互作用数据库整合在一起，用于评估受感染T淋巴细胞中铁超负荷调节的任何病毒辅因子。结果在这里，我们证明铁超负荷通过降低病毒RNA的水平下调HIV-1基因的表达。此外，我们发现铁超载调节许多病毒辅因子的表达。其中，REV辅因子eIF5A的下调可能与铁诱导的HIV-1基因表达的抑制有关。因此，我们证明了shRNA引起的eiF5A下调导致Nef水平显着降低，从而阻碍了HIV-1复制。结论我们的研究表明受铁代谢影响的HIV-1辅助因子代表抗逆转录病毒疗法的潜在靶标，并建议eIF5A作为药物开发的选择性靶标。我们发现铁过载调节许多病毒辅因子的表达。其中，REV辅因子eIF5A的下调可能与铁诱导的HIV-1基因表达的抑制有关。因此，我们证明了shRNA引起的eiF5A下调导致Nef水平显着降低，从而阻碍了HIV-1复制。结论我们的研究表明受铁代谢影响的HIV-1辅助因子代表抗逆转录病毒疗法的潜在靶标，并建议eIF5A作为药物开发的选择性靶标。我们发现铁过载调节许多病毒辅因子的表达。其中，REV辅因子eIF5A的下调可能与铁诱导的HIV-1基因表达的抑制有关。因此，我们证明了shRNA引起的eiF5A下调导致Nef水平显着降低，从而阻碍了HIV-1复制。结论我们的研究表明受铁代谢影响的HIV-1辅助因子代表抗逆转录病毒疗法的潜在靶标，并建议eIF5A作为药物开发的选择性靶标。