Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC 27599.
Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC 27599
Proc Natl Acad Sci U S A. 2018 Apr 10;115(15):E3408-E3415. doi: 10.1073/pnas.1801687115. Epub 2018 Mar 26.
We have adapted the eXcision Repair-sequencing (XR-seq) method to generate single-nucleotide resolution dynamic repair maps of UV-induced cyclobutane pyrimidine dimers and (6-4) pyrimidine-pyrimidone photoproducts in the genome. We find that these photoproducts are removed from the genome primarily by incisions 13-18 nucleotides 5' and 6-7 nucleotides 3' to the UV damage that generate 21- to 27-nt-long excision products. Analyses of the excision repair kinetics both in single genes and at the genome-wide level reveal strong transcription-coupled repair of the transcribed strand at early time points followed by predominantly nontranscribed strand repair at later stages. We have also characterized the excision repair level as a function of the transcription level. The availability of high-resolution and dynamic repair maps should aid in future repair and mutagenesis studies in this model organism.
我们已经采用了切除修复测序（XR-seq）方法，以生成在基因组中紫外线诱导的环丁烷嘧啶二聚体和（6-4）嘧啶-嘧啶酮光产物的单核苷酸分辨率动态修复图谱。我们发现，这些光产物主要通过在紫外线损伤处的 5'侧切割 13-18 个核苷酸和 3'侧切割 6-7 个核苷酸来从基因组中切除，从而产生 21-27 个核苷酸长的切除产物。在单个基因和全基因组水平上对切除修复动力学的分析表明，在早期阶段转录偶联修复转录链，随后在后期阶段主要修复非转录链。我们还将切除修复水平作为转录水平的函数进行了表征。高分辨率和动态修复图谱的可用性应该有助于未来在该模式生物中的修复和突变研究。
