PXD004782

PXD004782 is an original dataset announced via ProteomeXchange.

Title	Stable nuclear expression of ATP8 and ATP6 genes rescues a mtDNA Complex V null mutant
Description	We explore the possibility of re-engineering mitochondrial genes and expressing them from the nucleus as an approach to rescue defects arising from mitochondrial DNA mutations. We have used a patient cybrid cell line with a single point mutation in the overlap region of the ATP8 and ATP6 genes of the human mitochondrial genome. We show that these cells are null for the ATP8 protein, have significantly lowered ATP6 protein levels and no Complex V function. Nuclear expression of only the ATP8 gene with the ATP5G1 mitochondrial targeting sequence appended restored viability on Krebs cycle substrates and ATP synthesis capabilities but, failed to restore ATP hydrolysis and was insensitive to various inhibitors of oxidative phosphorylation. Co-expressing both ATP8 and ATP6 genes under similar conditions resulted in stable protein expression leading to successful integration into Complex V of the oxidative phosphorylation machinery. Tests for ATP hydrolysis / synthesis activity, oxygen consumption, glycolytic metabolism, and viability all indicate a significant functional rescue of the mutant phenotype following stable co-expression of ATP8 and ATP6. Thus we report the stable allotopic expression, import, and function of two mitochondria encoded genes, ATP8 and ATP6, resulting in the simultaneous rescue of the loss of both mitochondrial proteins.
HostingRepository	MassIVE
AnnounceDate	2016-10-05
AnnouncementXML	Submission_2016-10-05_09:34:33.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Birgit Schilling
SpeciesList	scientific name: Homo sapiens (human);
ModificationList	No PTMs are included in the dataset
Instrument	TripleTOF 6600; TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2016-08-12 02:24:10	ID requested
⏵ 1	2016-10-05 09:34:34	announced

Boominathan A, Vanhoozer S, Basisty N, Powers K, Crampton AL, Wang X, Friedricks N, Schilling B, Brand MD, O'Connor MS, Stable nuclear expression of ATP8 and ATP6 genes rescues a mtDNA Complex V null mutant. Nucleic Acids Res, 44(19):9342-9357(2016) [pubmed]

submitter keyword: SWATH, data independent acquisition, ATP6, mitochondria, Complex 5

Birgit Schilling
lab head
Birgit Schilling
contact affiliation	Buck Institute
contact email	bschilling@buckinstitute.org
dataset submitter

MassIVE dataset URI

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