Department of Chemistry , University of California , Riverside , California 92521-0403 , United States.
ACS Chem Biol. 2019 Mar 15;14(3):478-485. doi: 10.1021/acschembio.8b01126. Epub 2019 Feb 25.
Genomic integrity is constantly challenged by exposure to environmental and endogenous genotoxic agents. Reactive oxygen species (ROS) represent one of the most common types of DNA damaging agents. While ROS mainly induce single-nucleobase lesions, epimeric 2-deoxyribose lesions can also be induced upon hydrogen atom abstraction from the C1', C3', or C4' carbon and the subsequent incorrect chemical repair of the resulting carbon-centered radicals. Herein, we investigated the replicative bypass of the C1'- and C3'-epimeric lesions of the four 2'-deoxynucleosides in HEK293T human embryonic kidney epithelial cells. Our results revealed distinct bypass efficiencies and mutagenic properties of these two types of epimeric lesions. Replicative bypasses of all C1'-epimeric lesions except α-dA are mutagenic in HEK293T cells, and their mutagenic properties are further modulated by translesion synthesis (TLS) DNA polymerases. By contrast, none of the four C3'-epimeric lesions are mutagenic, and the replicative bypass of these lesions is not compromised upon depletion of polymerase η, ι, κ, or ζ. Together, our results provide important new knowledge about the cytotoxic and mutagenic properties of C1' and C3' epimeric lesions, and reveal the roles of TLS DNA polymerases in bypassing these lesions in human cells.
基因组完整性不断受到环境和内源性遗传毒性物质的挑战。活性氧(ROS)是最常见的 DNA 损伤剂之一。虽然 ROS 主要诱导单核苷酸损伤,但在从 C1'、C3'或 C4'碳原子上提取氢原子后,也可以诱导差向 2-脱氧核糖损伤,并且随后对产生的碳中心自由基进行不正确的化学修复。在此,我们研究了 HEK293T 人胚胎肾上皮细胞中四种 2'-脱氧核苷的 C1'和 C3'差向异构化损伤的复制旁路。我们的结果揭示了这两种类型的差向异构化损伤具有不同的旁路效率和诱变特性。除了 α-dA 之外,所有 C1'差向异构化损伤的复制旁路在 HEK293T 细胞中都是诱变的,并且它们的诱变特性进一步受到跨损伤合成(TLS)DNA 聚合酶的调节。相比之下,四种 C3'差向异构化损伤都没有诱变,并且当聚合酶 η、ι、κ 或 ζ 耗尽时,这些损伤的复制旁路不受影响。总之,我们的研究结果为 C1'和 C3'差向异构化损伤的细胞毒性和诱变特性提供了重要的新知识,并揭示了 TLS DNA 聚合酶在人类细胞中绕过这些损伤的作用。