Department of Biological Chemistry, The Institute of Life Sciences, The Hebrew University of Jerusalem, Edmond J. Safra Campus, Jerusalem, Israel.
Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan.
Biophys J. 2022 Aug 16;121(16):3126-3135. doi: 10.1016/j.bpj.2022.07.011. Epub 2022 Jul 9.
Transcription factor (TF) binding to genomic DNA elements constitutes one of the key mechanisms that regulates gene expression program in cells. Both consensus and nonconsensus DNA sequence elements influence the recognition specificity of TFs. Based on the analysis of experimentally determined c-Myc binding preferences to genomic DNA, here we statistically predict that certain repetitive, nonconsensus DNA symmetry elements can relatively reduce TF-DNA binding preferences. This is in contrast to a different set of repetitive, nonconsensus symmetry elements that can increase the strength of TF-DNA binding. Using c-Myc enhancer reporter system containing consensus motif flanked by nonconsensus sequences in embryonic stem cells, we directly demonstrate that the enrichment in such negatively regulating repetitive symmetry elements is sufficient to reduce the gene expression level compared with native genomic sequences. Negatively regulating repetitive symmetry elements around consensus c-Myc motif and DNA sequences containing consensus c-Myc motif flanked by entirely randomized sequences show similar expression baseline. A possible explanation for this observation is that rather than complete repression, negatively regulating repetitive symmetry elements play a regulatory role in fine-tuning the reduction of gene expression, most probably by binding TFs other than c-Myc.
转录因子(TF)与基因组 DNA 元件的结合是调节细胞中基因表达程序的关键机制之一。共识和非共识 DNA 序列元件都影响 TF 的识别特异性。基于对实验确定的 c-Myc 与基因组 DNA 结合偏好性的分析,我们在这里从统计学上预测,某些重复的、非共识的 DNA 对称元件可以相对降低 TF-DNA 的结合偏好性。这与另一组重复的、非共识的对称元件形成对比,后者可以增强 TF-DNA 的结合强度。使用含有非共识序列侧翼的共识基序的 c-Myc 增强子报告系统在胚胎干细胞中,我们直接证明,富含这种负调节重复对称元件足以降低基因表达水平,与天然基因组序列相比。围绕共识 c-Myc 基序的负调节重复对称元件和含有完全随机化序列侧翼的共识 c-Myc 基序的 DNA 序列显示出相似的表达基线。对这一观察结果的一种可能解释是,负调节重复对称元件不是完全抑制,而是在微调基因表达的降低方面发挥调节作用,很可能通过结合除 c-Myc 以外的 TF。