Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/118538
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Type: Journal article
Title: Capture of somatic mtDNA point mutations with severe effects on oxidative phosphorylation in synaptosome cybrid clones from human brain
Author: Mckenzie, M.
Chiotis, M.
Hroudová, J.
Lopez Sanchez, M.
Lim, S.
Cook, M.
Mckelvie, P.
Cotton, R.
Murphy, M.
St John, J.
Trounce, I.
Citation: Human Mutation, 2014; 35(12):1476-1484
Publisher: Wiley Online Library
Issue Date: 2014
ISSN: 1059-7794
1098-1004
Statement of
Responsibility: 
Matthew McKenzie, Maria Chiotis, Jana Hroudová, Maria I.G. Lopez Sanchez, Sze Chern Lim, Mark J. Cook, Penny McKelvie, Richard G. H. Cotton, Michael Murphy, Justin C. St John, Ian A. Trounce
Abstract: Mitochondrial DNA (mtDNA) is replicated throughout life in postmitotic cells, resulting in higher levels of somatic mutation than in nuclear genes. However, controversy remains as to the importance of low-level mtDNA somatic mutants in cancerous and normal human tissues. To capture somatic mtDNA mutations for functional analysis, we generated synaptosome cybrids from synaptic endings isolated from fresh hippocampus and cortex brain biopsies. We analyzed the whole mtDNA genome from 120 cybrid clones derived from four individual donors by chemical cleavage of mismatch and Sanger sequencing, scanning around two million base pairs. Seventeen different somatic point mutations were identified, including eight coding region mutations, four of which result in frameshifts. Examination of one cybrid clone with a novel m.2949_2953delCTATT mutation in MT-RNR2 (which encodes mitochondrial 16S rRNA) revealed a severe disruption of mtDNA-encoded protein translation. We also performed functional studies on a homoplasmic nonsense mutation in MT-ND1, previously reported in oncocytomas, and show that both ATP generation and the stability of oxidative phosphorylation complex I are disrupted. As the mtDNA remains locked against direct genetic manipulation, we demonstrate that the synaptosome cybrid approach can capture biologically relevant mtDNA mutants in vitro to study effects on mitochondrial respiratory chain function.
Keywords: Mitochondrial DNA; somatic mutation; synaptosome cybrid; oxidative phosphorylation
Rights: © 2014 WILEY PERIODICALS, INC.
RMID: 0030112578
DOI: 10.1002/humu.22694
Grant ID: http://purl.org/au-research/grants/nhmrc/1061472
Appears in Collections:Medicine publications

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