Surrallés Jordi, Ramírez María J, Marcos Ricard, Natarajan Adayapalam T, Mullenders Leon H F
Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10571-4. doi: 10.1073/pnas.162278199. Epub 2002 Jul 25.
A specialized nucleotide excision repair pathway known as transcription-coupled repair (TCR) counteracts the toxic effects of DNA damage in transcriptionally active genes. The clustering of active genes into gene-rich chromosomal domains predicts that the sites of TCR are unevenly distributed through the genome. To elucidate the genomic organization and chromosomal localization of TCR, we isolated DNA fragments encompassing TCR-mediated repair sites from UV-C irradiated xeroderma pigmentosum group C cells, which can only repair the transcribed strand of active genes. This DNA was used as a molecular probe to visualize TCR in normal metaphase spreads by reverse fluorescence in situ hybridization. Whereas DNA repair sites in normal human cells are evenly distributed through the genome, TCR is highly localized at specific chromosomal domains. Particularly, clusters of TCR sites were identified at early-replicating gene-rich bands and telomeric regions of several chromosomes. High gene-density chromosomes such as chromosome 19 and the GC-rich domains of several chromosomes (T bands) are preferential locations of TCR. Our results demonstrate that the intragenomic localization of TCR resembles the uneven distribution of the human transcriptome, CpG islands, and hyperacetylated histones, enforcing the basic link between DNA repair, transcription, and nuclear organization in a complex genome.
一种被称为转录偶联修复(TCR)的特殊核苷酸切除修复途径可抵消转录活跃基因中DNA损伤的毒性作用。活跃基因聚集在富含基因的染色体区域,这预示着TCR位点在基因组中的分布并不均匀。为了阐明TCR的基因组组织和染色体定位,我们从经紫外线C照射的C组着色性干皮病细胞中分离出包含TCR介导修复位点的DNA片段,这些细胞只能修复活跃基因的转录链。该DNA被用作分子探针,通过反向荧光原位杂交在正常中期染色体铺片中观察TCR。正常人类细胞中的DNA修复位点在基因组中均匀分布,而TCR则高度定位于特定的染色体区域。特别是,在几条染色体的早期复制富含基因的带区和端粒区域发现了TCR位点簇。高基因密度的染色体,如19号染色体以及几条染色体的富含GC的区域(T带)是TCR的优先定位位置。我们的结果表明,TCR在基因组内的定位类似于人类转录组、CpG岛和高度乙酰化组蛋白的不均匀分布,强化了复杂基因组中DNA修复、转录和核组织之间的基本联系。