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Rescue of a deficiency in ATP synthesis by transfer of MTATP6, a mitochondrial DNA-encoded gene, to the nucleus

Manfredi, Giovanni, Fu, Jin, Ojaimi, Joseline, Sadlock, James E., Kwong, Jennifer Q., Guy, John, Schon, Eric A.·2002·Nature Genetics
DOI 10.1038/ng851
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Abstract

A T-->G transversion at nt 8993 in mitochondrial DNA of MTATP6 (encoding ATPase 6 of complex V of the respiratory chain) causes impaired mitochondrial ATP synthesis in two related mitochondrial disorders: neuropathy, ataxia and retinitis pigmentosa and maternally inherited Leigh syndrome. To overcome the biochemical defect, we expressed wildtype ATPase 6 protein allotopically from nucleus-transfected constructs encoding an amino-terminal mitochondrial targeting signal appended to a recoded ATPase 6 gene (made compatible with the universal genetic code) that also contained a carboxy-terminal FLAG epitope tag. After transfection of human cells, the precursor polypeptide was expressed, imported into and processed within mitochondria, and incorporated into complex V. Allotopic expression of stably transfected constructs in cytoplasmic hybrids (cybrids) homoplasmic with respect to the 8993T-->G mutation showed a significantly improved recovery after growth in selective medium as well as a significant increase in ATP synthesis. This is the first successful demonstration of allotopic expression of an mtDNA-encoded polypeptide in mammalian cells and could form the basis of a genetic approach to treat a number of human mitochondrial disorders.

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  • The protease cleavage site of the ATP5G1 MTS has been determined, and the first five amino acids of mature ATP5G1 protein were included to facilitate processing.

    “The protease cleavage site of this MTS has been determined and, for this study, we also included the first five amino acids of the mature ATP5G1 protein to facilitate processing of the precursor [36].”

    From Codon optimization is an essential parameter for the efficient allotopic expression of mtDNA genes (2020) · cites this paper as [36]

  • Allotopic expression of ATP6 in patient cybrids with specific mutations such as mt.8993T>G or mt.8529G>A shows promise and can restore partial function.

    “Allotopic expression of ATP6 in patient cybrids with specific mutations such as the mt.8993T>G or (mt.8529G>A) shows promise and can restore partial function.27,29,40”

    From Exogenous expression of ATP8, a mitochondrial encoded protein, from the nucleus in vivo (2024) · cites this paper as [29]

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Metadata

Authors
Manfredi, G., Fu, J., Ojaimi, J., Sadlock, J. E., Kwong, J. Q., Guy, J., & Schon, E. A.
Venue
Nature Genetics
Publisher
Springer Science and Business Media LLC
Published
2002-02-25
Volume
30
Issue
4
Pages
394-399
ISSN
  • 1061-4036
  • 1546-1718
DOI
10.1038/ng851
Cited by
235
References
0
Language
en
Licence
www.springer.com/tdm (tdm)

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