DNA Polymerase η Promotes the Transcriptional Bypass of N2-Alkyl-2′-deoxyguanosine Adducts in Human Cells,Journal of the American Chemical Society

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DNA Polymerase η Promotes the Transcriptional Bypass of N2-Alkyl-2′-deoxyguanosine Adducts in Human Cells
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2021-09-23 , DOI: 10.1021/jacs.1c07374
Ying Tan ₁ , Su Guo ₁ , Jun Wu ₂ , Hua Du ₂ , Lin Li ₂ , Changjun You ₂ , Yinsheng Wang _{1,

2}

Affiliation

To cope with unrepaired DNA lesions, cells are equipped with DNA damage tolerance mechanisms, including translesion synthesis (TLS). While TLS polymerases are well documented in facilitating replication across damaged DNA templates, it remains unknown whether TLS polymerases participate in transcriptional bypass of DNA lesions in cells. Herein, we employed the competitive transcription and adduct bypass assay to examine the efficiencies and fidelities of transcription across N²-alkyl-2′-deoxyguanosine (N²-alkyl-dG, alkyl = methyl, ethyl, n-propyl, or n-butyl) lesions in HEK293T cells. We found that N²-alkyl-dG lesions strongly blocked transcription and elicited CC → AA tandem mutations in nascent transcripts, where adenosines were misincorporated opposite the lesions and their adjacent 5′ nucleoside. Additionally, genetic ablation of Pol η, but not Pol κ, Pol ι, or Pol ζ, conferred marked diminutions in the transcriptional bypass efficiencies of the N²-alkyl-dG lesions, which is exacerbated by codepletion of Rev1 in Pol η-deficient background. We also observed that the repair of N²-nBu-dG was not pronouncedly affected by genetic depletion of Pol η or Rev1. Hence, our results provided insights into transcriptional perturbations induced by N²-alkyl-dG lesions and expanded the biological functions of TLS DNA polymerases.

中文翻译：

DNA 聚合酶 η 促进人类细胞中 N2-烷基-2'-脱氧鸟苷加合物的转录旁路

为了应对未修复的 DNA 损伤，细胞配备了 DNA 损伤耐受机制，包括跨损伤合成 (TLS)。虽然 TLS 聚合酶在促进受损 DNA 模板的复制方面得到充分证明，但 TLS 聚合酶是否参与细胞中 DNA 损伤的转录旁路仍然未知。在这里，我们采用竞争性转录和加合物旁路测定来检查跨N ² -烷基-2'-脱氧鸟苷（N ² -烷基-dG，烷基 = 甲基、乙基、正丙基或n -丁基）HEK293T 细胞损伤。我们发现N ²-烷基-dG损伤强烈阻断转录并在新生转录物中引发CC→AA串联突变，其中腺苷与损伤及其相邻的5'核苷相对错入。此外，Pol η 的基因消融，但不是 Pol κ、Pol ι 或 Pol ζ，导致N ² -烷基-dG 病变的转录旁路效率显着降低，这因 Pol η 缺陷中 Rev1 的共同缺失而加剧背景。我们还观察到N ² - n Bu-dG 的修复不受 Pol η 或 Rev1 基因缺失的显着影响。因此，我们的结果提供了对N ²诱导的转录扰动的见解。-烷基-dG损伤并扩展了TLS DNA聚合酶的生物学功能。

更新日期：2021-10-06

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