Imaging assay to probe the role of telomere length shortening on telomere-gene interactions in single cells.
Imaging assay to probe the role of telomere length shortening on telomere-gene interactions in single cells.
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DOI：
10.1007/s00412-020-00747-4
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发表时间：
2021-03
期刊：
影响因子：
1.6
通讯作者：
Danuser G
中科院分区：
文献类型：
--
作者：
Zhang N;Li Y;Lai TP;Shay JW;Danuser G
端粒是覆盖染色体末端的重复非编码核苷酸序列（TTAGGGn）。随着年龄的增长，端粒的逐渐缩短与基因表达的变化有关，这是通过端粒位置效应（TPE）等模型实现的，这表明端粒对越来越远的基因的转录活性的干扰减少。TPE模型的一种修改，称为长距离端粒位置效应（TPE-OLD），解释了为什么距离端粒1-10 MB的一些基因仍然受到TPE的影响，但距离端粒更近的基因则没有。在这里，我们描述了一种成像方法，系统地检查发生TPE-OLD在单细胞水平。与现有方法相比，该管道允许快速分析数百至数千个细胞，这对于将TPE-OLD建立为可接受的基因表达调控机制是必要的。我们研究了两个人类基因，ISG 15和TERT，之前已经描述了TPE-OLD。对于这两个基因，我们发现与年轻细胞相比，老年细胞中同一染色体上的端粒相互作用较少;并且老年细胞中实验性延长的端粒挽救了两个基因的端粒相互作用水平。然而，从年轻细胞到老年细胞的年龄进程的相互作用的依赖性是不同的。ISG 15和TERT之间差异的一个模型可能与两个基因中显著不同的间质端粒序列排列有关。总之，这为端粒长度缩短在基因表达调控中的作用提供了强有力的理论基础。在线版本包含补充材料，可通过10.1007/s 00412 -020-00747-4获得。
Telomeres are repetitive non-coding nucleotide sequences (TTAGGGn) capping the ends of chromosomes. Progressive telomere shortening with increasing age has been associated with shifts in gene expression through models such as the telomere position effect (TPE), which suggests reduced interference of the telomere with transcriptional activity of increasingly more distant genes. A modification of the TPE model, referred to as Telomere Position Effects over Long Distance (TPE-OLD), explains why some genes 1–10 MB from a telomere are still affected by TPE, but genes closer to the telomere are not. Here, we describe an imaging approach to systematically examine the occurrence of TPE-OLD at the single cell level. Compared to existing methods, the pipeline allows rapid analysis of hundreds to thousands of cells, which is necessary to establish TPE-OLD as an acceptable mechanism of gene expression regulation. We examined two human genes, ISG15 and TERT, for which TPE-OLD has been described before. For both genes, we found less interaction with the telomere on the same chromosome in old cells compared to young cells; and experimentally elongated telomeres in old cells rescued the level of telomere interaction for both genes. However, the dependency of the interactions on the age progression from young to old cells varied. One model for the differences between ISG15 and TERT may relate to the markedly distinct interstitial telomeric sequence arrangement in the two genes. Overall, this provides a strong rationale for the role of telomere length shortening in the regulation of gene expression. The online version contains supplementary material available at 10.1007/s00412-020-00747-4.
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