Faculty of Life and Environmental Sciences, Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, Tsukuba, Ibaraki, Japan.
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
PLoS One. 2019 Feb 14;14(2):e0203099. doi: 10.1371/journal.pone.0203099. eCollection 2019.
Long-range associations between enhancers and their target gene promoters have been shown to play critical roles in executing genome function. Recent variations of chromosome capture technology have revealed a comprehensive view of intra- and interchromosomal contacts between specific genomic sites. The locus control region of the β-globin genes (β-LCR) is a super-enhancer that is capable of activating all of the β-like globin genes within the locus in cis through physical interaction by forming DNA loops. CTCF helps to mediate loop formation between LCR-HS5 and 3'HS1 in the human β-globin locus, in this way thought to contribute to the formation of a "chromatin hub". The β-globin locus is also in close physical proximity to other erythrocyte-specific genes located long distances away on the same chromosome. In this case, erythrocyte-specific genes gather together at a shared "transcription factory" for co-transcription. Theoretically, enhancers could also activate target gene promoters at the identical loci, yet on different chromosomes in trans, a phenomenon originally described as transvection in Drosophilla. Although close physical proximity has been reported for the β-LCR and the β-like globin genes when integrated at the mouse homologous loci in trans, their structural and functional interactions were found to be rare, possibly because of a lack of suitable regulatory elements that might facilitate such trans interactions. Therefore, we re-evaluated presumptive transvection-like enhancer-promoter communication by introducing CTCF binding sites and erythrocyte-specific transcription units into both LCR-enhancer and β-promoter alleles, each inserted into the mouse ROSA26 locus on separate chromosomes. Following cross-mating of mice to place the two mutant loci at the identical chromosomal position and into active chromation in trans, their transcriptional output was evaluated. The results demonstrate that there was no significant functional association between the LCR and the β-globin gene in trans even in this idealized experimental context.
长距离增强子与其靶基因启动子之间的关联已被证明在执行基因组功能方面起着关键作用。最近的染色体捕获技术的变化揭示了特定基因组位点之间的内染色体和染色体间接触的综合视图。β-珠蛋白基因(β-LCR)的基因调控区是一个超级增强子,能够通过形成 DNA 环,通过物理相互作用在顺式激活基因座内的所有β样珠蛋白基因。CTCF 有助于介导人类β-珠蛋白基因座中 LCR-HS5 和 3'HS1 之间的环形成,从而有助于形成“染色质枢纽”。β-珠蛋白基因座也与位于同一染色体上远距离的其他红细胞特异性基因密切物理接近。在这种情况下,红细胞特异性基因聚集在共享的“转录工厂”中进行共转录。理论上,增强子也可以在相同的基因座上激活靶基因启动子,但在不同的染色体上发生顺式作用,这种现象最初在果蝇中被描述为 transvection。尽管在小鼠同源基因座中转导时已经报道了β-LCR 和β样珠蛋白基因的紧密物理接近,但它们的结构和功能相互作用很少见,这可能是由于缺乏合适的调节元件来促进这种顺式相互作用。因此,我们通过引入 CTCF 结合位点和红细胞特异性转录单元到 LCR 增强子和β-启动子等位基因中,重新评估了假定的 transvection 样增强子-启动子通讯,每个基因都插入到小鼠 ROSA26 基因座的不同染色体上。在交叉交配小鼠将两个突变基因座置于相同的染色体位置并在顺式中激活染色质后,评估了它们的转录输出。结果表明,即使在这种理想化的实验背景下,LCR 和β-珠蛋白基因之间也没有显著的功能关联。