Department of Biology and Center for Evolutionary and Theoretical Immunology, University of New Mexico, Albuquerque, NM, United States of America.
Department of Biology, San Diego State University, San Diego, CA, United States of America.
PLoS One. 2022 Jul 21;17(7):e0271554. doi: 10.1371/journal.pone.0271554. eCollection 2022.
To gain understanding into the mechanisms of transcriptional activation of muscle genes, we sought to determine if genes targeted by the myogenic transcription factor Myocyte enhancer factor-2 (MEF2) were enriched for specific core promoter elements. We identified 330 known MEF2 target promoters in Drosophila, and analyzed them for for the presence and location of 17 known consensus promoter sequences. As a control, we also searched all Drosophila RNA polymerase II-dependent promoters for the same sequences. We found that promoter motifs were readily detected in the MEF2 target dataset, and that many of them were slightly enriched in frequency compared to the control dataset. A prominent sequence over-represented in the MEF2 target genes was NDM2, that appeared in over 50% of MEF2 target genes and was 2.5-fold over-represented in MEF2 targets compared to background. To test the functional significance of NDM2, we identified two promoters containing a single copy of NDM2 plus an upstream MEF2 site, and tested the activity of these promoters in vivo. Both the sticks and stones and Kahuli fragments showed strong skeletal myoblast-specific expression of a lacZ reporter in embryos. However, the timing and level of reporter expression was unaffected when the NDM2 site in either element was mutated. These studies identify variations in promoter architecture for a set of regulated genes compared to all RNA polymerase II-dependent genes, and underline the potential redundancy in the activities of some core promoter elements.
为了深入了解肌肉基因转录激活的机制,我们试图确定肌细胞增强因子 2(MEF2)的靶基因是否富含特定的核心启动子元件。我们在果蝇中鉴定了 330 个已知的 MEF2 靶启动子,并分析了它们是否存在 17 个已知的保守启动子序列。作为对照,我们还在所有依赖 RNA 聚合酶 II 的果蝇启动子中搜索了相同的序列。我们发现启动子基序在 MEF2 靶标数据集中很容易被检测到,并且与对照数据集相比,其中许多基序的频率略有富集。在 MEF2 靶基因中过度表达的一个突出序列是 NDM2,它出现在超过 50%的 MEF2 靶基因中,并且在 MEF2 靶基因中比背景过度表达 2.5 倍。为了测试 NDM2 的功能意义,我们鉴定了两个含有单个 NDM2 拷贝和一个上游 MEF2 位点的启动子,并在体内测试了这些启动子的活性。sticks and stones 和 Kahuli 片段在胚胎中的 lacZ 报告基因中均表现出强烈的骨骼肌成肌细胞特异性表达。然而,当两个元件中的任何一个 NDM2 位点发生突变时,报告基因的表达时间和水平均不受影响。这些研究确定了一组受调控基因的启动子结构与所有依赖 RNA 聚合酶 II 的基因相比存在差异,并强调了一些核心启动子元件的活性存在潜在冗余。