A genome-scale DNA repair RNAi screen identifies SPG48 as a novel gene associated with 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.
Słabicki
Mikołaj
M
Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany.
Theis
Mirko
M
Krastev
Dragomir B
DB
Samsonov
Sergey
S
Mundwiller
Emeline
E
Junqueira
Magno
M
Paszkowski-Rogacz
Maciej
M
Teyra
Joan
J
Heninger
Anne-Kristin
AK
Poser
Ina
I
Prieur
Fabienne
F
Truchetto
Jérémy
J
Confavreux
Christian
C
Marelli
Cécilia
C
Durr
Alexandra
A
Camdessanche
Jean Philippe
JP
Brice
Alexis
A
Shevchenko
Andrej
A
Pisabarro
M Teresa
MT
Stevanin
Giovanni
G
Buchholz
Frank
F
D004260:DNA Repair; D055785:Gene Knockdown Techniques; D015894:Genome, Human; D006801:Humans; D034622:RNA Interference; D011995:Recombination, Genetic; D015419:Spastic Paraplegia, Hereditary / Q000235:genetics*