Endogenous DNA damage at sites of terminated transcripts

• 成果类型: Article
• 署名作者: Liu, Jingjing; Perren, Jullian O.; Rogers, Cody M.; Nimer, Sadeieh; Wen, Alice X.; Halliday, Jennifer A.; Fitzgerald, Devon M.; Mei, Qian; Nehring, Ralf B.; Crum, Mary; Kozmin, Stanislav G.; Xia, Jun; Cooke, Matthew B.; Zhai, Yin; Bates, David; Li, Lei; Hastings, P. J.; Artsimovitch, Irina; Herman, Christophe; Sung, Patrick M.; Miller, Kyle M.; Rosenberg, Susan M.
• 署名单位: Baylor College of Medicine; Baylor College of Medicine; University of Texas System; University of Texas Austin; University of Texas System; University of Texas at San Antonio; Baylor College of Medicine; Baylor College of Medicine; University of Texas System; UTMD Anderson Cancer Center; University System of Ohio; Ohio State University; Emory University
• 刊物名称: Nature
• ISSN/ISSBN: 0028-1916
• DOI: 10.1038/s41586-024-08578-4
• 发表日期: 2025-04-03
• 关键词: escherichia-coli; genome instability; polymerase-iii; repair factors; fork reversal; replication; rna; recombination; mutations; mechanism

摘要

DNA damage promotes mutations that fuel cancer, ageing and neurodegenerative diseases1, 2-3, but surprisingly, the causes and types of damage remain largely unknown. There are three identified mechanisms that damage DNA during transcription: collision of RNA polymerase (RNAP) with the DNA-replication machinery head-on and co-directionally4, 5-6, and R-loop-induced DNA breakage7, 8, 9-10. Here we identify novel DNA damage reaction intermediates11,12 and uncover a fourth transcription-related source of DNA damage: endogenous DNA damage at sites of terminated transcripts. We engineered proteins to capture single-stranded DNA (ssDNA) ends with 3 ' polarity in bacterial and human cells. In Escherichia coli, spontaneous 3 '-ssDNA-end foci were unexpectedly frequent, at one or more per cell division, and arose via two identifiable pathways, both of which were dependent on DNA replication. A pathway associated with double-strand breaks was suppressed by overexpression of replicative DNA polymerase (pol) III, suggesting competition between pol III and DNA damage-promoting proteins. Mapping of recurrent 3 '-ssDNA-ends identified distinct 3 '-ssDNA-end-hotspots, mostly unrelated to double-strand breaks, next to the 5 '-CCTTTTTT transcription-terminator-like sequence. These 3 '-ssDNA-termini coincide with RNA 3 '-termini identified by DirectRNA sequencing13 or simultaneous 5 ' and 3 ' end RNA sequencing (SEnd-seq)14 and were prevented by a mutant RNAP that reads through terminators. Our findings reveal that transcription termination or pausing can promote DNA damage and subsequent genomic instability.