Heterologous mitochondrial DNA recombination in human cells

Marilena D'Aurelio, Carl D. Gajewski, Michael T. Lin, William M. Mauck, Leon Z. Shao, Giorgio Lenaz, Carlos T Moraes, Giovanni Manfredi

Research output: Contribution to journalArticle

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Abstract

Inter-molecular heterologous mitochondrial DNA (mtDNA) recombination is known to occur in yeast and plants. Nevertheless, its occurrence in human cells is still controversial. To address this issue we have fused two human cytoplasmic hybrid cell lines, each containing a distinct pathogenic mtDNA mutation and specific sets of genetic markers. In this hybrid model, we found direct evidence of recombination between these two mtDNA haplotypes. Recombinant mtDNA molecules in the hybrid cells were identified using three independent experimental approaches. First, recombinant molecules containing genetic markers from both parental alleles were demonstrated with restriction fragment length polymorphism of polymerase chain reaction products, by measuring the relative frequencies of each marker. Second, fragments of recombinant mtDNA were cloned and sequenced to identify the regions involved in the recombination events. Finally, recombinant molecules were demonstrated directly by Southern blot using appropriate combinations of polymorphic restriction sites and probes. This combined approach confirmed the existence of heterogeneous species of recombinant mtDNA molecules in the hybrid cells. These findings have important implications for mtDNA-related diseases, the interpretation of human evolution and population genetics and forensic analyses based on mtDNA genotyping.

Original languageEnglish
Pages (from-to)3171-3179
Number of pages9
JournalHuman Molecular Genetics
Volume13
Issue number24
DOIs
StatePublished - Dec 15 2004

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Mitochondrial DNA
Genetic Recombination
Recombinant DNA
Hybrid Cells
Genetic Markers
Medical Genetics
Population Genetics
Southern Blotting
Restriction Fragment Length Polymorphisms
Haplotypes
Yeasts
Alleles
Cell Line
Polymerase Chain Reaction
Mutation

ASJC Scopus subject areas

  • Genetics

Cite this

D'Aurelio, M., Gajewski, C. D., Lin, M. T., Mauck, W. M., Shao, L. Z., Lenaz, G., ... Manfredi, G. (2004). Heterologous mitochondrial DNA recombination in human cells. Human Molecular Genetics, 13(24), 3171-3179. https://doi.org/10.1093/hmg/ddh326

Heterologous mitochondrial DNA recombination in human cells. / D'Aurelio, Marilena; Gajewski, Carl D.; Lin, Michael T.; Mauck, William M.; Shao, Leon Z.; Lenaz, Giorgio; Moraes, Carlos T; Manfredi, Giovanni.

In: Human Molecular Genetics, Vol. 13, No. 24, 15.12.2004, p. 3171-3179.

Research output: Contribution to journalArticle

D'Aurelio, M, Gajewski, CD, Lin, MT, Mauck, WM, Shao, LZ, Lenaz, G, Moraes, CT & Manfredi, G 2004, 'Heterologous mitochondrial DNA recombination in human cells', Human Molecular Genetics, vol. 13, no. 24, pp. 3171-3179. https://doi.org/10.1093/hmg/ddh326
D'Aurelio M, Gajewski CD, Lin MT, Mauck WM, Shao LZ, Lenaz G et al. Heterologous mitochondrial DNA recombination in human cells. Human Molecular Genetics. 2004 Dec 15;13(24):3171-3179. https://doi.org/10.1093/hmg/ddh326
D'Aurelio, Marilena ; Gajewski, Carl D. ; Lin, Michael T. ; Mauck, William M. ; Shao, Leon Z. ; Lenaz, Giorgio ; Moraes, Carlos T ; Manfredi, Giovanni. / Heterologous mitochondrial DNA recombination in human cells. In: Human Molecular Genetics. 2004 ; Vol. 13, No. 24. pp. 3171-3179.
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