Huda Alajlan, Vlad-Stefan Raducanu, Yossef Lopez de Los Santos, Muhammad Tehseen, Hibah Alruwaili, Amer Al-Mazrou, Reem Mohammad, Monther Al-Alwan, Alfredo De Biasio, Jasmeen S Merzaban, Hamoud Al-Mousa, Samir M Hamdan, Anas M Alazami
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Here we report a homozygous LIG1 mutation (p.A624T), affecting a universally conserved residue, in a patient presenting with leukopenia, neutropenia, lymphopenia, pan-hypogammaglobulinemia, and diminished in vitro response to mitogen stimulation. Patient fibroblasts expressed normal levels of LIG1 protein but exhibited impaired growth, poor viability, high baseline levels of gamma-H2AX foci, and an enhanced susceptibility to DNA-damaging agents. The mutation reduced LIG1 activity by lowering its affinity for magnesium 2.5-fold. Remarkably, it also increased LIG1 fidelity > 50-fold against 3' end 8-Oxoguanine mismatches, exhibiting a marked reduction in its ability to process such nicks. This is expected to yield increased ss- and dsDNA breaks. Molecular dynamic simulations, and Residue Interaction Network studies, predicted an allosteric effect for this mutation on the protein loops associated with the LIG1 high-fidelity magnesium, as well as on DNA binding within the adenylation domain. 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引用次数: 0
摘要
细胞的生存能力和抗癌能力取决于其保持基因组完整性的能力,而当核酸磷酸二酯键被破坏时,基因组完整性就会受到严重损害。DNA 连接酶 1(LIG1)通过封闭 DNA 复制和修复过程中产生的单链缺口,在基因组维护中发挥着关键作用。该基因的常染色体隐性突变以前只在少数个体中出现过。在此,我们报告了一名患者的同基因 LIG1 突变(p.A624T),该突变影响了一个普遍保守的残基,患者表现为白细胞减少症、中性粒细胞减少症、淋巴细胞减少症、泛高丙种球蛋白血症以及对有丝分裂原刺激的体外反应减弱。患者的成纤维细胞表达正常水平的LIG1蛋白,但表现出生长受阻、存活率低、γ-H2AX病灶基线水平高以及对DNA损伤剂的敏感性增强。这种突变使 LIG1 对镁的亲和力降低了 2.5 倍,从而降低了其活性。值得注意的是,该突变还使 LIG1 对 3' 端 8-氧鸟嘌呤错配的保真度提高了 50 倍以上,明显降低了其处理此类缺口的能力。预计这将增加 ss 和 dsDNA 的断裂。分子动力学模拟和残基相互作用网络研究预测,这种突变会对与 LIG1 高保真镁相关的蛋白质环以及腺苷酸化结构域内的 DNA 结合产生异构效应。这些由单个突变引起的活性受抑和保真度增强的双重变化,强调了 LIG1 缺陷如何导致严重免疫疾病的机理。
Severe Combined Immunodeficiency from a Homozygous DNA Ligase 1 Mutant with Reduced Catalytic Activity but Increased Ligation Fidelity.
A cell's ability to survive and to evade cancer is contingent on its ability to retain genomic integrity, which can be seriously compromised when nucleic acid phosphodiester bonds are disrupted. DNA Ligase 1 (LIG1) plays a key role in genome maintenance by sealing single-stranded nicks that are produced during DNA replication and repair. Autosomal recessive mutations in a limited number of individuals have been previously described for this gene. Here we report a homozygous LIG1 mutation (p.A624T), affecting a universally conserved residue, in a patient presenting with leukopenia, neutropenia, lymphopenia, pan-hypogammaglobulinemia, and diminished in vitro response to mitogen stimulation. Patient fibroblasts expressed normal levels of LIG1 protein but exhibited impaired growth, poor viability, high baseline levels of gamma-H2AX foci, and an enhanced susceptibility to DNA-damaging agents. The mutation reduced LIG1 activity by lowering its affinity for magnesium 2.5-fold. Remarkably, it also increased LIG1 fidelity > 50-fold against 3' end 8-Oxoguanine mismatches, exhibiting a marked reduction in its ability to process such nicks. This is expected to yield increased ss- and dsDNA breaks. Molecular dynamic simulations, and Residue Interaction Network studies, predicted an allosteric effect for this mutation on the protein loops associated with the LIG1 high-fidelity magnesium, as well as on DNA binding within the adenylation domain. These dual alterations of suppressed activity and enhanced fidelity, arising from a single mutation, underscore the mechanistic picture of how a LIG1 defect can lead to severe immunological disease.
期刊介绍:
The Journal of Clinical Immunology publishes impactful papers in the realm of human immunology, delving into the diagnosis, pathogenesis, prognosis, or treatment of human diseases. The journal places particular emphasis on primary immunodeficiencies and related diseases, encompassing inborn errors of immunity in a broad sense, their underlying genotypes, and diverse phenotypes. These phenotypes include infection, malignancy, allergy, auto-inflammation, and autoimmunity. We welcome a broad spectrum of studies in this domain, spanning genetic discovery, clinical description, immunologic assessment, diagnostic approaches, prognosis evaluation, and treatment interventions. Case reports are considered if they are genuinely original and accompanied by a concise review of the relevant medical literature, illustrating how the novel case study advances the field. The instructions to authors provide detailed guidance on the four categories of papers accepted by the journal.