Salinero Alicia C, Emerson Simey, Cormier Tayla C, Yin John, Morse Randall H, Curcio M Joan
Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, NY, United States.
Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, NY, United States.
Front Mol Biosci. 2022 Jul 1;9:896215. doi: 10.3389/fmolb.2022.896215. eCollection 2022.
The Ty1 retrotransposon family is maintained in a functional but dormant state by its host, . Several hundred and genes encoding co-factors and restrictors of Ty1 retromobility, respectively, have been identified. Well-characterized examples include and , encoding subunits of the Mediator transcriptional co-activator complex; control of retromobility by Med3 and Med15 requires the Ty1 promoter in the U3 region of the long terminal repeat. To characterize the U3-dependence of other Ty1 regulators, we screened a library of 188 known and mutants for altered retromobility of Ty1 expressed from the strong, TATA-less promoter or the weak, TATA-containing U3 promoter. Two classes of genes, each including both s and s, were identified. The first class comprising 82 genes that regulated Ty1 retromobility independently of U3 is enriched for genes that restrict the G1 phase of the cell cycle and those involved in transcriptional elongation and mRNA catabolism. The second class of 51 genes regulated retromobility of Ty1 driven only from the U3 promoter. Nineteen U3-dependent regulators (U3DRs) also controlled retromobility of Ty1 driven by the weak, TATA-less promoter, suggesting reliance on the low activity of U3. Thirty-one U3DRs failed to modulate P -Ty1 retromobility, suggesting dependence on the architecture of U3. To further investigate the U3-dependency of Ty1 regulators, we developed a novel fluorescence-based assay to monitor expression of p22-Gag, a restriction factor expressed from the internal Ty1i promoter. Many U3DRs had minimal effects on levels of Ty1 RNA, Ty1i RNA or p22-Gag. These findings uncover a role for the Ty1 promoter in integrating signals from diverse host factors to modulate Ty1 RNA biogenesis or fate.
Ty1逆转录转座子家族由其宿主维持在一种功能但休眠的状态。已经分别鉴定出数百个编码Ty1逆转移动性的辅助因子和限制因子的基因。特征明确的例子包括编码中介体转录共激活复合物亚基的基因和基因;Med3和Med15对逆转移动性的控制需要长末端重复序列U3区域中的Ty1启动子。为了表征其他Ty1调节因子对U3的依赖性,我们筛选了一个包含188个已知基因和基因的突变体文库,以检测从强的、无TATA的启动子或弱的、含TATA的U3启动子表达的Ty1的逆转移动性变化。鉴定出了两类基因,每类都包括基因和基因。第一类包括82个独立于U3调节Ty1逆转移动性的基因,这些基因富含限制细胞周期G1期的基因以及参与转录延伸和mRNA分解代谢的基因。第二类51个基因仅调节从U3启动子驱动的Ty1的逆转移动性。19个U3依赖性调节因子(U3DRs)也控制了由弱的、无TATA的启动子驱动的Ty1的逆转移动性,这表明依赖于U3的低活性。31个U3DRs未能调节P -Ty1的逆转移动性,这表明依赖于U3的结构。为了进一步研究Ty1调节因子对U3的依赖性,我们开发了一种基于荧光标记的新方法来监测从内部Ty1i启动子表达的限制因子p22-Gag的表达。许多U3DRs对Ty1 RNA、Ty1i RNA或p22-Gag的水平影响极小。这些发现揭示了Ty1启动子在整合来自不同宿主因子的信号以调节Ty1 RNA生物合成或命运方面的作用。