A Genome-Scale DNA Repair RNAi Screen Identifies SPG48 as a Novel Gene Associated with Hereditary Spastic Paraplegia

作者：Slabicki Mikolaj^*; Theis Mirko; Krastev Dragomir B; Samsonov Sergey; Mundwiller Emeline; Junqueira Magno; Paszkowski Rogacz Maciej; Teyra Joan; Heninger Anne Kristin; Poser Ina; Prieur Fabienne; Truchetto Jeremy; Confavreux Christian; Marelli Cecilia; Durr Alexandra; Camdessanche Jean Philippe; Brice Alexis; Shevchenko Andrej; Pisabarro M Teresa; Stevanin Giovanni; Buchholz Frank

来源：PLoS Biology, 2010, 8(6): e1000408.

DOI：10.1371/journal.pbio.1000408

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.

• 出版日期2010-6