Assignment of two mitochondrially synthesized polypeptides to human mitochondrial DNA and their use in the study of intracellular mitochondrial interaction.

Oliver, Noëlynn; Wallace, Douglas C.

doi:10.1128/mcb.2.1.30

Cited by 83 publications

(41 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The mitochondrial translation patterns observed in the absence of CAP in 143B cells on one side and in VA2B and CAP23 cells on the other were identical except for the exclusive presence of the ND3 normal gene product in 143B cells and for a greatly reduced amount of the ND3 polypeptide and a predominant amount of the variant ND3' product in VA2B and CAP23 cells. The coexistence of a small amount of the ND3' variant polypeptide with a prevalent normal ND3 product had been reported previously for HeLa cells (35). Also to be noticed is the fact that the labeling of the mitochondrial translation products in CAP23 cells was significantly reduced compared with that in either 143B cells (by 25%) or VA2B cells (by 39%), in agreement with earlier observations (50 (Fig.…”

Section: Resultssupporting

confidence: 82%

“…An interesting observation is the relatively high level of labeling of the mitochondrial translation products (a60% of the level in 143B) and rateOf o2consump-tion (p30% of the level in 143B) in the parental line 2SD, which had the highest mtDNA content (o3. indicated by the experiments discussed in the previous section, was at variance with previously reported findings (35). These findings had been interpreted to suggest that HT1080 mtDNA, carrying a 16S rRNA gene mutation that conferred CAP resistance of mitochondrial protein synthesis on the cells, allowed the synthesis of a HeLa cell mtDNA-encoded variant ND3 polypeptide (originally named MV2 and subsequently renamed ND3' on the basis of the functional identification of the ND3 gene [9]) in the presence of CAP after introduction by cybrid or hybrid fusion into HeLa cells.…”

Section: Resultscontrasting

confidence: 56%

“…However, in both of the above-mentioned cases, the results could not distinguish between the possibility that the presence of normal and deleted mtDNAs in the same mitochondrion resulted from fusion of two homoplasmic mitochondria, one containing only deleted mtDNA and the other containing only wild-type mtDNA, and the preexistence of both mtDNAs in the same mitochondrion from the time of the mutation event. Evidence which had been interpreted to suggest an intermixing and cooperation of gene products of mutant and wild-type mtDNAs originally carried in distinct organelles had been reported in an earlier study involving hybrids or cybrids between chloramphenicol (CAP)-sensitive (CAPS) human cells and CAP-resistant (CAPr) cells or cytoplasts derived therefrom, respectively (35).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Complementation of mutant and wild-type human mitochondrial DNAs coexisting since the mutation event and lack of complementation of DNAs introduced separately into a cell within distinct organelles.

1994

View full text Add to dashboard Cite

The rules that govern complementation of mutant and wild-type mitochondrial genomes in human cells were investigated under different experimental conditions. Among mitochondrial transformants derived from an individual affected by the MERRF (myoclonus epilepsy associated with ragged red fibers) encephalomyopathy and carrying in heteroplasmic form the mitochondrial tRNALYS mutation associated with that syndrome, normal protein synthesis and respiration was observed when the wild-type mitochondrial DNA exceeded 10% of the total complement. In these transformants, the protective effect of wild-type mitochondrial DNA was shown to involve interactions of the mutant and wild-type gene products. Very different results were obtained in experiments in which two mitochondrial DNAs carrying nonallelic disease-causing mutations were sequentially introduced within distinct organelles into the same human mitochondrial DNA-less (p°) cell. In transformants exhibiting different ratios of the two genomes, no evidence of cooperation between their products was observed, even 3 months after the introduction of the second mutation. These results pointed to the phenotypic independence of the two genomes. A similar conclusion was reached in experiments in which mitochondria carrying a chloramphenicol resistance-inducing mitochondrial DNA mutation were introduced into chloramphenicol-sensitive cells. A plausible interpretation of the different results obtained in the latter two sets of experiments, compared with the complementation behavior observed in the heteroplasmic MERRF transformants, is that in the latter, the mutant and wild-type genomes coexisted in the same organelles from the time of the mutation. This would imply that the way in which mitochondrial DNA is sorted among different organelles plays a fundamental role in determining the oxidative-phosphorylation phenotype in mammalian cells. These results have significant implications for mitochondrial genetics and for studies on the transmission and therapy of mitochondrial DNA-linked diseases.Despite the large number of studies on the expression of artificially produced mitochondrial DNA (mtDNA) mutations and the phenotype of somatic cell hybrids and cybrids, information on mitochondrial genome interactions in mammalian cells is still very limited. The dependence of such interactions on the distribution of the mtDNA molecules among the mitochondria, which changes continuously throughout division and, possibly, fusion of the organelles, and the difficulty of investigating mtDNA sorting in a cell account in part for the present lack of understanding of how much the mitochondrial genomes interact in mammalian cells. The recent discovery of a variety of mtDNA mutations associated with diseases in humans (47) and the growing evidence of accumulation of mtDNA mutations with aging in somatic tissues (12-14, 19, 43) have raised a number of questions about the occurrence and frequency of intermixing of mutant and wild-type mtDNA and/or their products within a cell and the role that co...

show abstract

Section: Resultssupporting

confidence: 82%

Section: Resultscontrasting

confidence: 56%

mentioning

confidence: 99%

See 1 more Smart Citation

Complementation of mutant and wild-type human mitochondrial DNAs coexisting since the mutation event and lack of complementation of DNAs introduced separately into a cell within distinct organelles.

1994

View full text Add to dashboard Cite

show abstract

“…Using somatic cell fusion techniques, the occurrence of interaction of the mammalian mitochondrial genetic system was proved in heteroplasmic cells by translational complementation of mitochondrial rRNA between mitochondria with chloramphenicolsensitive and -resistant mtDNAs (7,8), and by translational complementation or competition of mitochondrial tRNAs between mitochondria with wild-type and pathogenic deletion mutant mtDNAs (8,9), although extensive recombination was not observed in heteroplasmic cells with rat mtDNAs or rat and mouse mtDNAs (10). Subsequently, we found rapid penetration of HeLa mtDNA and/or its products into mitochondria of mtDNA-less ( 0 ) HeLa cells in cybrids isolated by the fusion of enucleated HeLa cells with 0 HeLa cells (11).…”

mentioning

confidence: 99%

Transcomplementation between Different Types of Respiration-deficient Mitochondria with Different Pathogenic Mutant Mitochondrial DNAs

Takai

Isobe

Hayashi

1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

“…However, this mtDNA transfer system is problematic in that the mtDNA donor cells must be resistant to chloramphenicol for selective isolation of cells with exogenously transferred mtDNA (cybrids) (5,6). Moreover, chloramphenicol selection and subsequent cultivation in the presence of chloramphenicol could not completely remove endogenous wild-type (chloramphenicol-sensitive) mtDNA in the host cells (7,8). Therefore, the influence of the remaining host-cell mtDNA on the expression of the phenotypes cannot be excluded completely.…”

mentioning

confidence: 99%

Isolation of Mitochondrial DNA-less Mouse Cell Lines and Their Application for Trapping Mouse Synaptosomal Mitochondrial DNA with Deletion Mutations

Inoue

Ito

Takai

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

For isolation of mouse mtDNA-less ( 0 ) cell lines, we searched for various antimitochondrial drugs that were expected to decrease the mtDNA content and found that treatment with ditercalinium, an antitumor bis-intercalating agent, was extremely effective for completely excluding mtDNA in all the mouse cell lines we tested. The resulting 0 mouse cells were successfully used for trapping the mtDNA of living nerve cells into dividing cultured cells by fusion of the 0 cells with mouse brain synaptosomes, which represent synaptic endings isolated from nerve cells. With neuronal mtDNA obtained, all of the cybrid clones restored mitochondrial translation activity similarly regardless of whether the mtDNA was derived from young or aged mice, thus at least suggesting that defects in mitochondrial genomes are not involved in the age-associated mitochondrial dysfunction observed in the brain of aged mice. Furthermore, we could trap a very small amount of a common 5823-base pair deletion mutant mtDNA (⌬mtDNA 5823 ) that was detectable by polymerase chain reaction in the cybrid clones. As the amount of mutant mtDNA with large scale deletions was expected to increase during prolonged cultivation of the cybrids, these cells should be available for establishment of mice containing the deletion mutant mtDNA.

show abstract

Assignment of two mitochondrially synthesized polypeptides to human mitochondrial DNA and their use in the study of intracellular mitochondrial interaction.

Cited by 83 publications

References 49 publications

Complementation of mutant and wild-type human mitochondrial DNAs coexisting since the mutation event and lack of complementation of DNAs introduced separately into a cell within distinct organelles.

Complementation of mutant and wild-type human mitochondrial DNAs coexisting since the mutation event and lack of complementation of DNAs introduced separately into a cell within distinct organelles.

Transcomplementation between Different Types of Respiration-deficient Mitochondria with Different Pathogenic Mutant Mitochondrial DNAs

Isolation of Mitochondrial DNA-less Mouse Cell Lines and Their Application for Trapping Mouse Synaptosomal Mitochondrial DNA with Deletion Mutations

Contact Info

Product

Resources

About