BACKGROUND Prototype foamy virus (PFV) is a complex and unique retrovirus with the longest genome among the retroviruses and is used as a vector for gene therapies. The viral Tas protein transactivates the viral long terminal repeat promoter and is required for viral replication. We have utilized RNA sequencing to identify and characterize the long-noncoding RNA NONHSAG000101 (lnc-NONH), which markedly increases in PFV-infected cells. However, little is known about the function of lnc-NONH. OBJECTIVES We aim to explore the role of lnc-NONH during PFV infection. METHODS To assess the lnc-NONH role during PFV infection, the siRNAs were used to silence the lnc-NONH expression. The microRNA (miRNA) mimic and inhibitor were employed to explore the function of lnc-NONH-related miRNA miR-34c-5p. Quantitative real-time polymerase chain reaction assay and Western blotting were applied to measure the mRNA and protein levels of PFV transactivator Tas. Luciferase assay was used to determine the transcriptional activity of the PFV unique internal promoter (IP). RESULTS lnc-NONH promotes the expression of PFV Tas and miR-34c-5p. The interaction between lnc-NONH and miR-34c-5p enhances the transcriptional activity of the PFV IP. CONCLUSIONS In the current study, we report a novel mechanism for the lnc-NONH-mediated upregulation of Tas expression. Our findings contribute to the understanding of regulatory network of Tas expression and PFV replication.

中文翻译：

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长非编码RNA lnc-NONH通过上调miR-34c-5p和Tas蛋白的表达来增强泡沫型病毒的早期转录。

背景技术原型泡沫病毒（PFV）是复杂且独特的逆转录病毒，具有逆转录病毒中基因组最长的基因组，并且被用作基因疗法的载体。病毒Tas蛋白可激活病毒长末端重复启动子，是病毒复制所必需的。我们已利用RNA测序来鉴定和表征长非编码RNA NONHSAG000101（lnc-NONH），该蛋白在PFV感染的细胞中明显增加。但是，对于lnc-NONH的功能知之甚少。目的我们旨在探讨lnc-NONH在PFV感染中的作用。方法为了评估lnc-NONH在PFV感染过程中的作用，使用siRNA沉默lnc-NONH表达。使用microRNA（miRNA）模拟物和抑制剂来研究lnc-NONH相关miRNA miR-34c-5p的功能。实时定量聚合酶链反应测定和蛋白质印迹法用于测量PFV反式激活因子Tas的mRNA和蛋白水平。萤光素酶测定法用于测定PFV独特内部启动子（IP）的转录活性。结果lnc-NONH促进PFV Tas和miR-34c-5p的表达。lnc-NONH与miR-34c-5p之间的相互作用增强了PFV IP的转录活性。结论在当前研究中，我们报告了lnc-NONH介导的Tas表达上调的新机制。我们的发现有助于理解Tas表达和PFV复制的调控网络。萤光素酶测定法用于测定PFV独特内部启动子（IP）的转录活性。结果lnc-NONH促进了PFV Tas和miR-34c-5p的表达。lnc-NONH与miR-34c-5p之间的相互作用增强了PFV IP的转录活性。结论在当前研究中，我们报告了lnc-NONH介导的Tas表达上调的新机制。我们的发现有助于理解Tas表达和PFV复制的调控网络。萤光素酶测定法用于测定PFV独特内部启动子（IP）的转录活性。结果lnc-NONH促进PFV Tas和miR-34c-5p的表达。lnc-NONH与miR-34c-5p之间的相互作用增强了PFV IP的转录活性。结论在当前研究中，我们报告了lnc-NONH介导的Tas表达上调的新机制。我们的发现有助于理解Tas表达和PFV复制的调控网络。