title: High-throughput, pooled sequencing identifies mutations in NUBPL and FOXRED1 in human complex I deficiency.
abstract: Discovering the molecular basis of mitochondrial respiratory chain disease is challenging given the large number of both mitochondrial and nuclear genes that are involved. We report a strategy of focused candidate gene prediction, high-throughput sequencing and experimental validation to uncover the molecular basis of mitochondrial complex I disorders. We created seven pools of DNA from a cohort of 103 cases and 42 healthy controls and then performed deep sequencing of 103 candidate genes to identify 151 rare variants that were predicted to affect protein function. We established genetic diagnoses in 13 of 60 previously unsolved cases using confirmatory experiments, including cDNA complementation to show that mutations in NUBPL and FOXRED1 can cause complex I deficiency. Our study illustrates how large-scale sequencing, coupled with functional prediction and experimental validation, can be used to identify causal mutations in individual cases.
authors.affiliation: Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Calvo
authors.forename: Elena J
authors.initials: EJ
authors.lastname: Tucker
authors.forename: Alison G
authors.initials: AG
authors.lastname: Compton
authors.forename: Denise M
authors.initials: DM
authors.lastname: Kirby
authors.forename: Gabriel
authors.initials: G
authors.lastname: Crawford
authors.forename: Noel P
authors.initials: NP
authors.lastname: Burtt
authors.forename: Manuel
authors.initials: M
authors.lastname: Rivas
authors.forename: Candace
authors.initials: C
authors.lastname: Guiducci
authors.forename: Damien L
authors.initials: DL
authors.lastname: Bruno
authors.forename: Olga A
authors.initials: OA
authors.lastname: Goldberger
authors.forename: Michelle C
authors.initials: MC
authors.lastname: Redman
authors.forename: Esko
authors.initials: E
authors.lastname: Wiltshire
authors.forename: Callum J
authors.initials: CJ
authors.lastname: Wilson
authors.forename: David
authors.initials: D
authors.lastname: Altshuler
authors.forename: Stacey B
authors.initials: SB
authors.lastname: Gabriel
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Daly
authors.forename: David R
authors.initials: DR
authors.lastname: Thorburn
authors.forename: Vamsi K
authors.initials: VK
authors.lastname: Mootha
mesh_terms: D015153:Blotting, Western; D016022:Case-Control Studies; D042967:Electron Transport Complex I / Q000235:genetics*; D018628:Gene Dosage; D056726:Genetic Association Studies; D006801:Humans; D028361:Mitochondrial Diseases / Q000235:genetics*; D024101:Mitochondrial Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D042967:Electron Transport Complex I
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