Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, Mexico City, Mexico.
Consejo Nacional de Ciencia y Tecnología, Mexico City, Mexico.
Front Cell Infect Microbiol. 2018 Jul 12;8:214. doi: 10.3389/fcimb.2018.00214. eCollection 2018.
The protozoan parasite is exposed to reactive oxygen and nitric oxide species that have the potential to damage its genome. harbors enzymes involved in DNA repair pathways like Base and Nucleotide Excision Repair. The majority of DNA repairs pathways converge in their final step in which a DNA ligase seals the DNA nicks. In contrast to other eukaryotes, the genome of encodes only one DNA ligase (EhDNAligI), suggesting that this ligase is involved in both DNA replication and DNA repair. Therefore, the aim of this work was to characterize EhDNAligI, its ligation fidelity and its ability to ligate opposite DNA mismatches and oxidative DNA lesions, and to study its expression changes and localization during and after recovery from UV and HO treatment. We found that EhDNAligI is a high-fidelity DNA ligase on canonical substrates and is able to discriminate erroneous base-pairing opposite DNA lesions. EhDNAligI expression decreases after DNA damage induced by UV and HO treatments, but it was upregulated during recovery time. Upon oxidative DNA damage, EhDNAligI relocates into the nucleus where it co-localizes with EhPCNA and the 8-oxoG adduct. The appearance and disappearance of 8-oxoG during and after both treatments suggest that DNA damaged was efficiently repaired because the mainly NER and BER components are expressed in this parasite and some of them were modulated after DNA insults. All these data disclose the relevance of EhDNAligI as a specialized and unique ligase in that may be involved in DNA repair of the 8-oxoG lesions.
原生动物寄生虫 暴露于具有潜在基因组损伤能力的活性氧和一氧化氮物种中。 拥有参与 DNA 修复途径的酶,如碱基和核苷酸切除修复。大多数 DNA 修复途径在其最后一步汇聚,其中 DNA 连接酶封闭 DNA 缺口。与其他真核生物不同, 的基因组仅编码一种 DNA 连接酶(EhDNAligI),这表明该连接酶参与 DNA 复制和 DNA 修复。因此,这项工作的目的是表征 EhDNAligI、其连接保真度以及连接 DNA 错配和氧化 DNA 损伤的能力,并研究其在 UV 和 HO 处理期间和之后的表达变化和定位。我们发现 EhDNAligI 是一种高保真 DNA 连接酶,可在规范的底物上区分错误的碱基配对 DNA 损伤。EhDNAligI 的表达在 UV 和 HO 处理引起的 DNA 损伤后降低,但在恢复期间上调。在氧化 DNA 损伤后,EhDNAligI 重新定位到细胞核中,在那里它与 EhPCNA 和 8-oxoG 加合物共定位。在这两种处理过程中和之后 8-oxoG 的出现和消失表明 DNA 损伤被有效地修复,因为主要的 NER 和 BER 成分在这种寄生虫中表达,并且一些成分在 DNA 损伤后被调节。所有这些数据都揭示了 EhDNAligI 在 中的重要性,它可能是一种参与 8-oxoG 损伤 DNA 修复的特殊和独特的连接酶。