A Genome-Scale DNA Repair RNAi Screen Identifies SPG48 as a Novel Gene Associated with Hereditary Spastic Paraplegia
Open Access
- 31 May 2010
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLoS Biology
- Vol. 8 (6), e1000408
- https://doi.org/10.1371/journal.pbio.1000408
Abstract
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.Keywords
This publication has 63 references indexed in Scilit:
- CYP7B1 mutations in pure and complex forms of hereditary spastic paraplegia type 5Brain, 2009
- Separating the Wheat from the Chaff: Unbiased Filtering of Background Tandem Mass Spectra Improves Protein IdentificationJournal of Proteome Research, 2008
- Chromatin Central: towards the comparative proteome by accurate mapping of the yeast proteomic environmentGenome Biology, 2008
- Exon deletions of SPG4 are a frequent cause of hereditary spastic paraplegiaJournal of Medical Genetics, 2007
- Mutations in SPG11, encoding spatacsin, are a major cause of spastic paraplegia with thin corpus callosumNature Genetics, 2007
- In-gel digestion for mass spectrometric characterization of proteins and proteomesNature Protocols, 2006
- A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recoveryNature, 2005
- DNA Repair, Genome Stability, and AgingCell, 2005
- The DNA Double-Strand Break Repair Gene hMRE11 Is Mutated in Individuals with an Ataxia-Telangiectasia-like DisorderCell, 1999
- CLASSIFICATION OF THE HEREDITARY ATAXIAS AND PARAPLEGIASThe Lancet, 1983