Stimpson Kaitlin M, Sullivan Lori L, Kuo Molly E, Sullivan Beth A
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America; Institute for Genome Sciences & Policy, Duke University, Durham, North Carolina, United States of America.
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America.
PLoS One. 2014 Mar 24;9(3):e92432. doi: 10.1371/journal.pone.0092432. eCollection 2014.
The short arms of the ten acrocentric human chromosomes share several repetitive DNAs, including ribosomal RNA genes (rDNA). The rDNA arrays correspond to nucleolar organizing regions that coalesce each cell cycle to form the nucleolus. Telomere disruption by expressing a mutant version of telomere binding protein TRF2 (dnTRF2) causes non-random acrocentric fusions, as well as large-scale nucleolar defects. The mechanisms responsible for acrocentric chromosome sensitivity to dysfunctional telomeres are unclear. In this study, we show that TRF2 normally associates with the nucleolus and rDNA. However, when telomeres are crippled by dnTRF2 or RNAi knockdown of TRF2, gross nucleolar and chromosomal changes occur. We used the controllable dnTRF2 system to precisely dissect the timing and progression of nucleolar and chromosomal instability induced by telomere dysfunction, demonstrating that nucleolar changes precede the DNA damage and morphological changes that occur at acrocentric short arms. The rDNA repeat arrays on the short arms decondense, and are coated by RNA polymerase I transcription binding factor UBF, physically linking acrocentrics to one another as they become fusogenic. These results highlight the importance of telomere function in nucleolar stability and structural integrity of acrocentric chromosomes, particularly the rDNA arrays. Telomeric stress is widely accepted to cause DNA damage at chromosome ends, but our findings suggest that it also disrupts chromosome structure beyond the telomere region, specifically within the rDNA arrays located on acrocentric chromosomes. These results have relevance for Robertsonian translocation formation in humans and mechanisms by which acrocentric-acrocentric fusions are promoted by DNA damage and repair.
人类十条近端着丝粒染色体的短臂共享几种重复DNA,包括核糖体RNA基因(rDNA)。rDNA阵列对应于核仁组织区,在每个细胞周期中聚集形成核仁。通过表达端粒结合蛋白TRF2的突变体版本(dnTRF2)破坏端粒会导致非随机的近端着丝粒融合以及大规模的核仁缺陷。近端着丝粒染色体对功能失调的端粒敏感的机制尚不清楚。在本研究中,我们表明TRF2通常与核仁及rDNA相关联。然而,当端粒因dnTRF2或TRF2的RNA干扰敲低而受损时,会发生明显的核仁及染色体变化。我们使用可控的dnTRF2系统精确剖析了端粒功能障碍诱导的核仁及染色体不稳定的时间和进程,证明核仁变化先于近端着丝粒短臂处发生的DNA损伤和形态变化。短臂上的rDNA重复阵列解聚,并被RNA聚合酶I转录结合因子UBF覆盖,当近端着丝粒变得具有融合性时,它们在物理上相互连接。这些结果突出了端粒功能在近端着丝粒染色体的核仁稳定性和结构完整性中的重要性,特别是rDNA阵列。端粒应激被广泛认为会导致染色体末端的DNA损伤,但我们的研究结果表明,它还会破坏端粒区域之外的染色体结构,特别是位于近端着丝粒染色体上的rDNA阵列内。这些结果与人类罗伯逊易位的形成以及DNA损伤和修复促进近端着丝粒 - 近端着丝粒融合的机制相关。