Injection of mitochondria into human cells leads to a rapid replacement of the endogenous mitochondrial DNA.
Isolated human mitochondria containing a mitochondrial DNA (mtDNA) coded chloramphenicol resistance marker were injected into cells from two different human sensitive cell lines, 143BTK- and HT1080-6TG, which had been partially depleted of their mtDNA by ethidium bromide treatment. On the basis of the available evidence concerning the tolerance of introduced volumes into mammalian cells, it is estimated that, on the average, less than one mitochondrion was introduced into each cell. Under selective conditions, the mitochondria became established in the recipient cells with a frequency greater than 2-3 x 10(-3). An analysis of multiple mtDNA and nuclear DNA polymorphisms revealed a rapid replacement of the resident mtDNA by the exogenous mtDNA. Six to ten weeks after microinjection, this replacement was complete in all but one of the HT1080-6TG transformants, and nearly complete in the majority of the 143BTK- transformants. The quantitative behavior of the mtDNA of the transformants at very early stages of selection strongly suggests that intracellular mtDNA selection played a crucial role in this replacement, with significant implications for mitochondrial genetics.
King
M P
MP
Division of Biology, California Institute of Technology, Pasadena 91125.
Attardi
G
G
D002460:Cell Line; D002702:Chloramphenicol Resistance; D003001:Cloning, Molecular; D004272:DNA, Mitochondrial / Q000235:genetics*; D004996:Ethidium / Q000494:pharmacology; D006801:Humans; D008845:Microinjections; D008928:Mitochondria / Q000637:transplantation*; D009154:Mutation; D014162:Transfection
D004272:DNA, Mitochondrial; D004996:Ethidium