Baur Joseph A, Shay Jerry W, Wright Woodring E
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9039, USA.
Chromosoma. 2004 Feb;112(5):240-6. doi: 10.1007/s00412-003-0269-x. Epub 2004 Jan 20.
Subtelomeric reporter genes in human cells are silenced in a telomere length-dependent manner. Here we show that a subtelomeric reporter gene is expressed in only a subpopulation of cells within a clone and that this heterogeneity is generated by switching between expression states. We observed frequent reversion from the silenced state back to active expression. This process was more prominent for subtelomeric transgenes; however, we also observed cases of spontaneous reversion in control clones bearing the reporter at an internal site. We additionally show that treatment of these cells with 5-bromodeoxyuridine results in strong activation of the transgene. Although similar findings have been reported previously in mouse cells, this is, to our knowledge, the first direct observation of ongoing fluctuations in transcription in clonal populations of human cells. Our results suggest that this mechanism, as opposed to progressive silencing or a delayed fixing of expression states, accounts for the variegation in expression observed for subtelomeric transgenes in human cells. These data imply that telomere shortening during human aging could lead to stochastic activation of subtelomeric genes.
人类细胞中的亚端粒报告基因以端粒长度依赖的方式被沉默。在此我们表明,亚端粒报告基因仅在克隆内的一部分细胞亚群中表达,并且这种异质性是由表达状态之间的转换产生的。我们观察到从沉默状态频繁回复到活性表达。这个过程对于亚端粒转基因更为显著;然而,我们也在内部位点携带报告基因的对照克隆中观察到自发回复的情况。我们还表明,用5-溴脱氧尿苷处理这些细胞会导致转基因的强烈激活。尽管之前在小鼠细胞中已报道过类似的发现,但据我们所知,这是首次对人类细胞克隆群体中转录的持续波动进行直接观察。我们的结果表明,与渐进性沉默或表达状态的延迟固定相反,这种机制解释了人类细胞中亚端粒转基因表达的斑驳现象。这些数据意味着人类衰老过程中端粒缩短可能导致亚端粒基因的随机激活。