Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Institute of Biology Leiden, Leiden University, Leiden, The Netherlands.
PLoS Genet. 2020 Apr 24;16(4):e1008759. doi: 10.1371/journal.pgen.1008759. eCollection 2020 Apr.
Bases within DNA are frequently damaged, producing obstacles to efficient and accurate DNA replication by replicative polymerases. Translesion synthesis (TLS) polymerases, via their ability to catalyze nucleotide additions to growing DNA chains across DNA lesions, promote replication of damaged DNA, thus preventing checkpoint activation, genome instability and cell death. In this study, we used C. elegans to determine the contribution of TLS activity on long-term stability of an animal genome. We monitored and compared the types of mutations that accumulate in REV1, REV3, POLH1 and POLK deficient animals that were grown under unchallenged conditions. We also addressed redundancies in TLS activity by combining all deficiencies. Remarkably, animals that are deficient for all Y-family polymerases as well as animals that have lost all TLS activity are viable and produce progeny, demonstrating that TLS is not essential for animal life. Whole genome sequencing analyses, however, reveal that TLS is needed to prevent genomic scars from accumulating. These scars, which are the product of polymerase theta-mediated end joining (TMEJ), are found overrepresented at guanine bases, consistent with TLS suppressing DNA double-strand breaks (DSBs) from occurring at replication-blocking guanine adducts. We found that in C. elegans, TLS across spontaneous damage is predominantly error free and anti-clastogenic, and thus ensures preservation of genetic information.
DNA 中的碱基经常受到损伤,这会对复制酶进行有效且准确的 DNA 复制造成阻碍。跨损伤合成(TLS)聚合酶能够在 DNA 损伤处催化延伸 DNA 链的核苷酸添加,从而促进受损 DNA 的复制,防止检验点激活、基因组不稳定和细胞死亡。在这项研究中,我们利用秀丽隐杆线虫来确定 TLS 活性对动物基因组长期稳定性的贡献。我们监测并比较了在无挑战条件下生长的 REV1、REV3、POLH1 和 POLK 缺失动物中积累的突变类型。我们还通过组合所有缺失来解决 TLS 活性的冗余问题。值得注意的是,所有 Y 家族聚合酶缺失的动物以及失去所有 TLS 活性的动物都是有活力的,并能产生后代,这表明 TLS 对于动物的生命并不是必需的。然而,全基因组测序分析表明,TLS 对于防止基因组疤痕的积累是必要的。这些疤痕是聚合酶θ介导的末端连接(TMEJ)的产物,在鸟嘌呤碱基处过度表达,这与 TLS 抑制复制受阻的鸟嘌呤加合物处发生 DNA 双链断裂(DSBs)一致。我们发现,在秀丽隐杆线虫中,TLS 对自发损伤主要是无错误且抗突变的,从而确保遗传信息的保存。