Research Paper

Pharmaceutical Research

, 28:2871

First online:

DNA Delivery to Mitochondria: Sequence Specificity and Energy Enhancement

  • Noha IbrahimAffiliated withInstitut de Biologie Moléculaire des Plantes, CNRS and Université de StrasbourgMitochondrial Research Group, Institute for Aging & Health Medical School, Newcastle University
  • , Hirokazu HandaAffiliated withPlant Genome Research Unit, National Institute of Agrobiological Sciences
  • , Anne CossetAffiliated withInstitut de Biologie Moléculaire des Plantes, CNRS and Université de Strasbourg
  • , Milana KoulintchenkoAffiliated withInstitut de Biologie Moléculaire des Plantes, CNRS and Université de StrasbourgMitochondrial Research Group, Institute for Aging & Health Medical School, Newcastle UniversitySiberian Institute of Plant Physiology and Biochemistry, Russian Academy of Sciences
  • , Yuri KonstantinovAffiliated withSiberian Institute of Plant Physiology and Biochemistry, Russian Academy of Sciences
  • , Robert N. LightowlersAffiliated withMitochondrial Research Group, Institute for Aging & Health Medical School, Newcastle University
  • , André DietrichAffiliated withInstitut de Biologie Moléculaire des Plantes, CNRS and Université de Strasbourg Email author 
  • , Frédérique Weber-LotfiAffiliated withInstitut de Biologie Moléculaire des Plantes, CNRS and Université de Strasbourg

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ABSTRACT

Purpose

Mitochondria are competent for DNA uptake in vitro, a mechanism which may support delivery of therapeutic DNA to complement organelle DNA mutations. We document here key aspects of the DNA import process, so as to further lay the ground for mitochondrial transfection in intact cells.

Methods

We developed DNA import assays with isolated mitochondria from different organisms, using DNA substrates of various sequences and sizes. Further import experiments investigated the possible role of ATP and protein phosphorylation in the uptake process. The fate of adenine nucleotides and the formation of phosphorylated proteins were analyzed.

Results

We demonstrate that the efficiency of mitochondrial uptake depends on the sequence of the DNA to be translocated. The process becomes sequence-selective for large DNA substrates. Assays run with a natural mitochondrial plasmid identified sequence elements which promote organellar uptake. ATP enhances DNA import and allows tight integration of the exogenous DNA into mitochondrial nucleoids. ATP hydrolysis has to occur during the DNA uptake process and might trigger phosphorylation of co-factors.

Conclusions

Our data contribute critical information to optimize DNA delivery into mitochondria and open the prospect of targeting whole mitochondrial genomes or complex constructs into mammalian organelles in vitro and in vivo.

KEY WORDS

DNA import mitochondrial disease mitochondrial plasmid organelle transfection protein phosphorylation