Corrigendum: 5-Formylcytosine mediated DNA-protein cross-links block DNA replication and induce mutations in human cells.
Corrigendum: 5-Formylcytosine mediated DNA-protein cross-links block DNA replication and induce mutations in human cells.
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勘误表：5-甲酰胞嘧啶介导的 DNA-蛋白质交联可阻断 DNA 复制并诱导人类细胞突变。
DOI：
10.1093/nar/gky809
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发表时间：
2018
影响因子：
14.9
通讯作者：
Tretyakova,NataliaY
中科院分区：
文献类型：
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作者：
Ji,Shaofei;Fu,Iwen;Naldiga,Spandana;Shao,Hongzhao;Basu,AshisK;Broyde,Suse;Tretyakova,NataliaY
5-甲酰胞嘧啶（5 fC）是通过泰特蛋白介导的5-甲基-dC氧化引入的表观遗传DNA修饰。我们最近报道了5 fC与活细胞中的组蛋白形成可逆的DNA-蛋白缀合物（DPC）（Jiet al.（2017年）Angew.化学国际版，56：14130-14134）。我们现在检查5 fC介导的DPC对DNA复制的影响。在人跨损伤合成DNA聚合酶η和κ的存在下，对含有5 fC和含赖氨酸的蛋白质和肽之间的位点特异性DPC的合成DNA双链体进行引物延伸实验。我们发现含有组蛋白H2 A或H4的DPC完全抑制DNA复制，但当蛋白质进行蛋白水解消化时，复制阻断被去除。与11-mer或31-mer肽的交联被两种聚合酶以易错方式绕过，诱导靶向C→T转换和-1缺失。当含有5 fC-肽交联的质粒在人胚肾（HEK）293 T细胞中复制时，观察到类似类型的突变。对与人聚合酶η和κ结合的11聚体肽-dC交联的分子模拟显示，肽在DNA大沟侧很好地拟合，并且修饰的dC通过聚合酶活性位点中的摆动碱基配对与进入的dATP形成稳定的错配。
5-Formylcytosine (5fC) is an epigenetic DNA modification introduced via TET protein-mediated oxidation of 5-methyl-dC. We recently reported that 5fC form reversible DNA–protein conjugates (DPCs) with histone proteins in living cells (Jiet al.(2017)Angew. Chem. Int. Ed., 56:14130–14134). We now examined the effects of 5fC mediated DPCs on DNA replication. Synthetic DNA duplexes containing site-specific DPCs between 5fC and lysine-containing proteins and peptides were subjected to primer extension experiments in the presence of human translesion synthesis DNA polymerases η and κ. We found that DPCs containing histones H2A or H4 completely inhibited DNA replication, but the replication block was removed when the proteins were subjected to proteolytic digestion. Cross-links to 11-mer or 31-mer peptides were bypassed by both polymerases in an error-prone manner, inducing targeted C→T transitions and –1 deletions. Similar types of mutations were observed when plasmids containing 5fC-peptide cross-links were replicated in human embryonic kidney (HEK) 293T cells. Molecular simulations of the 11-mer peptide-dC cross-links bound to human polymerases η and κ revealed that the peptide fits well on the DNA major groove side, and the modified dC forms a stable mismatch with incoming dATP via wobble base pairing in the polymerase active site.