Highly efficient DNA synthesis in isolated mitochondria from rat liver

We have developed a highly efficient DNA-synthesizing system with isolated intact rat liver mitochondria. The ATP requirements for this in organello DNA synthesis are provided by endogenous synthesis in the presence of exogenous ADP and an oxidizable substrate. In this system, mitochondrial DNA synt...

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Bibliographic Details
Published in:Nucleic acids research 1994-05, Vol.22 (10), p.1861-1865
Main Authors: Enríquez, José A., Ramos, Javier, Pérez-Martos, Acisclo, López-Pérez, Manuel J., Montoya, Julio
Format: Article
Language:English
Subjects:
Adenosine Diphosphate - metabolism
Animals
Biological and medical sciences
Cations, Divalent - metabolism
Cell Fractionation
DNA Replication - physiology
DNA, Mitochondrial - analysis
DNA, Mitochondrial - biosynthesis
Fundamental and applied biological sciences. Psychology
Magnesium - metabolism
Male
Manganese - metabolism
Mitochondria, Liver - metabolism
Molecular and cellular biology
Molecular genetics
Rats
Rats, Wistar
Replication
RNA Probes
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Summary:We have developed a highly efficient DNA-synthesizing system with isolated intact rat liver mitochondria. The ATP requirements for this in organello DNA synthesis are provided by endogenous synthesis in the presence of exogenous ADP and an oxidizable substrate. In this system, mitochondrial DNA synthesis strikingly proceeds at a constant rate for about 5 h at 37°C. Gel electrophoresis, hybridization and restriction enzyme analyses show that intact mitochondria synthesize nucleic acids with a size of 16.5 kb, that correspond to mitochondrial DNA, and that both DNA strands are replicated. This in organello DNA synthesis requires the supply of dNTPs and decreases at high ADP concentration in the incubation medium.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/22.10.1861