A genome-scale DNA repair RNAi screen identifies SPG48 as a novel gene associated with hereditary spastic paraplegia.
A genome-scale DNA repair RNAi screen identifies SPG48 as a novel gene associated with hereditary spastic paraplegia.
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DOI：
10.1371/journal.pbio.1000408
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发表时间：
2010-06-29
期刊：
影响因子：
9.8
通讯作者：
Buchholz F
中科院分区：
文献类型：
--
作者：
Słabicki M;Theis M;Krastev DB;Samsonov S;Mundwiller E;Junqueira M;Paszkowski-Rogacz M;Teyra J;Heninger AK;Poser I;Prieur F;Truchetto J;Confavreux C;Marelli C;Durr A;Camdessanche JP;Brice A;Shevchenko A;Pisabarro MT;Stevanin G;Buchholz F
我们在全基因组双链断裂DNA修复RNAi筛选中发现了一个新的基因，并显示其与神经系统疾病遗传性痉挛性截瘫有关。DNA修复对于维持基因组完整性至关重要，并且在DNA修复中起作用的基因在各种人类疾病中经常发生突变。通过同源重组的修复通常恢复原始DNA序列而不引入突变，并且已经鉴定了同源重组DNA双链断裂修复（HR-DSBR）所需的许多基因。然而，这一重要的DNA修复途径在哺乳动物细胞中的系统分析还没有报道。在这里，我们描述了一个基因组规模的核糖核酸内切酶制备的短干扰RNA（esiRNA）屏幕参与DNA双链断裂修复的基因。我们报告了61个影响HR-DSBR频率的基因，并详细描述了其中一个降低HR-DSBR频率的基因。我们发现，基因KIAA 0415编码一个假定的解旋酶，与SPG 11和SPG 15，遗传性痉挛性截瘫（HSP）突变的两种蛋白质相互作用。我们鉴定了HSP患者的突变，发现KIAA 0415/SPG 48是一种新的HSP相关基因，并表明KIAA 0415/SPG 48突变细胞系对DNA损伤药物更敏感。我们提出了哺乳动物细胞中HR-DSBR的第一个基因组规模的调查，提供了一个数据集，应该加速发现在DNA修复和相关医疗条件中发挥作用的新基因。形成新型蛋白质复合物的蛋白质是高效HR-DSBR所需的，并且在患有HSP的患者中发生突变，这一发现表明HSP与DNA修复之间存在联系。我们身体中的所有细胞都必须科普其DNA的许多损伤。细胞使用一组基因来修复DNA并保持基因组的完整性。考虑到完整基因组的重要性，在许多人类疾病中具有DNA修复作用的基因发生突变也就不足为奇了。在这里，我们展示了人类细胞中基因组规模DNA修复筛选的结果，并发现了61个在这一过程中具有潜在作用的基因。我们详细研究了以前未知的基因（KIAA 0415/SPG 48），并证明了其有效的DNA双链断裂修复的重要性。进一步的分析揭示了SPG 48基因在一些遗传性痉挛性截瘫（HSP）患者中的突变。我们发现SPG 48与其他HSP蛋白质发生物理相互作用，并且患者细胞对DNA损伤药物敏感。我们的数据表明HSP和DNA修复之间存在联系，我们建议将来对HSP患者进行KIAA 0415/SPG 48突变筛查。
We have identified a novel gene in a genome-wide, double-strand break DNA repair RNAi screen and show that is involved in the neurological disease hereditary spastic paraplegia. DNA repair is essential to maintain genome integrity, and genes with roles in DNA repair are frequently mutated in a variety of human diseases. Repair via homologous recombination typically restores the original DNA sequence without introducing mutations, and a number of genes that are required for homologous recombination DNA double-strand break repair (HR-DSBR) have been identified. However, a systematic analysis of this important DNA repair pathway in mammalian cells has not been reported. Here, we describe a genome-scale endoribonuclease-prepared short interfering RNA (esiRNA) screen for genes involved in DNA double strand break repair. We report 61 genes that influenced the frequency of HR-DSBR and characterize in detail one of the genes that decreased the frequency of HR-DSBR. We show that the gene KIAA0415 encodes a putative helicase that interacts with SPG11 and SPG15, two proteins mutated in hereditary spastic paraplegia (HSP). We identify mutations in HSP patients, discovering KIAA0415/SPG48 as a novel HSP-associated gene, and show that a KIAA0415/SPG48 mutant cell line is more sensitive to DNA damaging drugs. We present the first genome-scale survey of HR-DSBR in mammalian cells providing a dataset that should accelerate the discovery of novel genes with roles in DNA repair and associated medical conditions. The discovery that proteins forming a novel protein complex are required for efficient HR-DSBR and are mutated in patients suffering from HSP suggests a link between HSP and DNA repair. All cells in our bodies have to cope with numerous lesions to their DNA. Cells use a battery of genes to repair DNA and maintain genome integrity. Given the importance of an intact genome, it is not surprising that genes with roles in DNA repair are mutated in many human diseases. Here, we present the results of a genome-scale DNA repair screen in human cells and discover 61 genes that have a potential role in this process. We studied in detail a previously uncharacterized gene (KIAA0415/SPG48) and demonstrated its importance for efficient DNA double strand break repair. Further analyses revealed mutations in the SPG48 gene in some patients with hereditary spastic paraplegia (HSP). We showed that SPG48 physically interacts with other HSP proteins and that patient cells are sensitive to DNA damaging drugs. Our data suggest a link between HSP and DNA repair and we propose that HSP patients should be screened for KIAA0415/SPG48 mutations in the future.
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DOI：
10.1073/pnas.0409861102
发表时间：
2005-02-15
影响因子：
11.1
作者：
Kittler, R;Pelletier, L;Buchholz, F
通讯作者：
Buchholz, F
影响因子：
4.4
作者：
Junqueira, Magno;Spirin, Victor;Shevchenko, Andrej
通讯作者：
Shevchenko, Andrej
影响因子：
14.5
作者：
Goizet, Cyril;Boukhris, Amir;Stevanin, Giovanni
通讯作者：
Stevanin, Giovanni
影响因子：
64.5
作者：
Doil, Carsten;Mailand, Niels;Lukas, Claudia
通讯作者：
Lukas, Claudia
DOI：
10.1042/bj20090942
发表时间：
2009-09-25
期刊：
The Biochemical journal
影响因子：
--
作者：
Hartlerode AJ;Scully R
通讯作者：
Scully R