Novel PNKP mutations associated with reduced DNA single-strand break repair and severe microcephaly, seizures, and developmental delay.
Novel PNKP mutations associated with reduced DNA single-strand break repair and severe microcephaly, seizures, and developmental delay.
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DOI：
10.1002/mgg3.2295
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发表时间：
2024-01
影响因子：
2
通讯作者：
中科院分区：
文献类型：
--
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小头畸形伴早发性癫痫发作（MCSZ）是一种由DNA链断裂修复蛋白、多核苷酸激酶3′-磷酸酶（PNKP）的致病性变体引起的神经发育障碍。我们使用全基因组测序和桑格测序来鉴定致病变异体，然后使用小基因测定、蛋白质印迹、碱性彗星试验、γ H2 AX和ADP-核糖免疫荧光。在这里，我们描述了一个病人与复合杂合子变异PNKP，包括错义变异的DNA磷酸酶域（T323 M）和一个新的剪接受体位点的DNA激酶域内的变异，我们显示导致外显子跳跃。我们发现，来自患者的原代成纤维细胞表现出PNKP蛋白水平大大降低，DNA单链断裂修复率降低，证实突变的PNKP等位基因功能失调。所提供的数据表明，检测到的复合杂合变体导致PNKP蛋白水平降低，这会影响氧化和TOP 1诱导的单链断裂的修复，并且最有可能导致该患者的MCSZ。小头畸形伴早发性癫痫发作（MSCZ）是一种由DNA链断裂修复蛋白、多核苷酸激酶3 '磷酸酶（PNKP）的致病性变体引起的神经发育障碍。在这里，我们描述了一个病人的复合杂合变异PNKP，一个错义变异，和一个新的剪接受体位点变异，导致外显子跳跃。我们发现，来自患者的原代成纤维细胞表现出PNKP蛋白水平大大降低，DNA单链断裂修复率降低，证实突变的PNKP等位基因功能失调。
Microcephaly with early‐onset seizures (MCSZ) is a neurodevelopmental disorder caused by pathogenic variants in the DNA strand break repair protein, polynucleotide kinase 3′‐phosphatase (PNKP). We have used whole genome sequencing and Sanger sequencing to identify disease‐causing variants, followed by a minigene assay, Western blotting, alkaline comet assay, γH2AX, and ADP‐ribose immunofluorescence. Here, we describe a patient with compound heterozygous variants in PNKP, including a missense variant in the DNA phosphatase domain (T323M) and a novel splice acceptor site variant within the DNA kinase domain that we show leads to exon skipping. We show that primary fibroblasts derived from the patient exhibit greatly reduced levels of PNKP protein and reduced rates of DNA single‐strand break repair, confirming that the mutated PNKP alleles are dysfunctional. The data presented show that the detected compound heterozygous variants result in reduced levels of PNKP protein, which affect the repair of both oxidative and TOP1‐induced single‐strand breaks, and most likely causes MCSZ in this patient. Microcephaly with early‐onset seizures (MSCZ) is a neurodevelopmental disorder caused by pathogenic variants in the DNA strand break repair protein, polynucleotide kinase 3′‐phophatase (PNKP). Here, we describe a patient with compound heterozygous variants in PNKP, one missense variant, and a novel splice acceptor site variant which leads to exon skipping. We show that primary fibroblasts derived from the patient exhibit greatly reduced levels of PNKP protein and reduced rates of DNA single‐strand break repair, confirming that the mutated PNKP alleles are dysfunctional.
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影响因子：
64.5
作者：
Loizou, JI;El-Khamisy, SF;Caldecott, KW
通讯作者：
Caldecott, KW
影响因子：
9.8
作者：
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通讯作者：
Guerreiro, Rita
影响因子：
0.4
作者：
Rudenskaya, Galina E.;Marakhonov, Andrey, V;Konovalov, Fedor A.
通讯作者：
Konovalov, Fedor A.
影响因子：
4.8
作者：
Karimi-Busheri, F;Daly, G;Weinfeld, M
通讯作者：
Weinfeld, M
影响因子：
3.8
作者：
Chalasani SL;Kawale AS;Akopiants K;Yu Y;Fanta M;Weinfeld M;Povirk LF
通讯作者：
Povirk LF