Mitochondrial genome expression in a mutant strain of<i>D.subobscura</i>, an animal model for large scale mtDNA deletion

Beziat, Florence; Morel, Frédéric; Volz-Lingenhol, Andrea; Paul, Noel Saint; Alziari, Serge

doi:10.1093/nar/21.3.387

Cited by 30 publications

(32 citation statements)

References 19 publications

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“…Total DNA was obtained from 50 -60 heads, thoraces, or abdomens according to the method described by F. Béziat et al (13).…”

Section: Total Dna Preparationmentioning

confidence: 99%

“…Heteroplasmy was determined by Southern blotting of DNA fragments obtained after digesting DNA with MspI and hybridization with the COIII probe labeled as described (13). The signals were analyzed by densitometry.…”

Section: Measurement Of Heteroplasmymentioning

confidence: 99%

“…The studied RNAs are 12 S RNA and COIII (genes not involved in deletion), cytochrome b, and ND1, ND5, ND4-4L (genes involved in deletion), using the probes described in Ref. 13.…”

Section: Measurement Of Rna Contentmentioning

confidence: 99%

“…The first investigations, performed in the whole adult fly (13), showed a strong heteroplasmic tendency in the deleted molecules; this affected 78% of the mitochondrial genomes. mtDNA cell content (per nuclear genome) showed a 50% increase in the mutant strain compared with the D. subobscura wild strain.…”

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confidence: 99%

“…On the other hand, the transcript concentrations of the concerned genes showed a decrease, although this was less than expected and differed according to the genes (from 55% for ND1 to 35% for cytochrome b). Furthermore, detection of the fusion transcript (13) showed that both types of genomes were expressed. Maximum respiratory complex activities were identical in the wild type and mutant strains for complex IV.…”

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Biochemical and Molecular Consequences of Massive Mitochondrial Gene Loss in Different Tissues of a Mutant Strain of Drosophila subobscura

Beziat

Touraille

Debise

et al. 1997

Journal of Biological Chemistry

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In the studied mutant strain of Drosophila subobscura, 78% of the mitochondrial genomes lost >30% of the coding region by deletion. The mutations was genetically stable. Despite this massive loss of mitochondrial genes, the mutant did not seem to be affected. Distribution of the two genome types, cell levels of mitochondrial DNA, steady-state concentrations of the mitochondrial gene transcripts, mitochondrial enzymatic activities, and ATP synthesis capacities were measured in the head, thorax, and abdomen fractions of the mutant strain in comparison with a wild type strain. Results indicate that the deleted genomes are detected in all fractions but to a lesser extent in the male and female abdomen. In all fractions, there is a 50% increase in cellular mitochondrial DNA content. Although there is a decrease in steady-state concentrations of mitochondrial transcripts of genes affected by deletion, this is smaller than expected. The variations in mitochondrial biochemical activities in the different fractions of the wild strain are upheld in the mutant strain. Activity of complex I (involved in mutation) nevertheless shows a decrease in all fractions; activity of complex III (likewise involved) shows little or no change; finally, mitochondrial ATP synthesis capacity is identical to that observed in the wild strain. This latter finding possibly accounts for the lack of phenotype. This mutant is a good model for studying mitochondrial genome alterations and the role of the nuclear genome in these phenomena.Various alterations in the mitochondrial genome have been correlated with severe human pathology (1-3). Among the most substantial of these alterations, deletions, which account for a small fraction of the described cases, have always been detected at the heteroplasmic state (4 -6). Once the proportion of deleted genomes reaches a critical threshold, these deletions have often very pejorative effects on mitochondrial function (7,8). Such mutations are generally sporadic, although dominant autosomal factors have been observed (9 -11). A locus has been localized on chromosome 10, but no gene has yet been identified.In the heteroplasmic state, the mutant Drosophila subobscura strain studied in our laboratory presents a substantial mtDNA 1 deletion (30% of the coding region) (12). Contrary to clinical findings (1), this mutation remains stable from one generation to the next. It does not seem to affect the mutant (apparent absence of phenotype) in terms of reproduction (number of embryos, emergence of larvae), life span, or flight capacities. This strain thus offers a good model for studying possible biochemical and molecular consequences of severe mitochondrial genome alteration and the responses that enable the mutant to survive.The first investigations, performed in the whole adult fly (13), showed a strong heteroplasmic tendency in the deleted molecules; this affected 78% of the mitochondrial genomes. mtDNA cell content (per nuclear genome) showed a 50% increase in the mutant strain compared with the D. subobscura ...

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“…Total DNA was obtained from 50 -60 heads, thoraces, or abdomens according to the method described by F. Béziat et al (13).…”

Section: Total Dna Preparationmentioning

confidence: 99%

Section: Measurement Of Heteroplasmymentioning

confidence: 99%

“…The studied RNAs are 12 S RNA and COIII (genes not involved in deletion), cytochrome b, and ND1, ND5, ND4-4L (genes involved in deletion), using the probes described in Ref. 13.…”

Section: Measurement Of Rna Contentmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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