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Nucleic Acids Research, 1978, Vol. 5, No. 1 285-295
© 1978

Articles

Enzymatic synthesis of DNA complementary to mitochondrial mRNA via reverse transcription

L. Frolova*, S. Arsenyan*, T. Avdonina*, V. Gaitskhoki, O. Kisselev, S. Neifach and L. Kisselev*,+

*Institute of Molecular Biology, USSR Academy of Science Vavilov str., 32, Moscow 117312 Institute of Experimental Medicine, USSR Academy of Medical Sciences Leningrad, USSR

+To whom to address correspondence

Received November 25, 1977. The poly(A)-containing mitochondrial mRNAs of rat liver were tested for their ability to serve as templates for the DNA synthesis by means of reverse transcription in the presence of the oligo(dT) primer and the RNA-directed DNA-polymerase from avian myeloblastoais virus. The mt-mRNA does not support the DNA synthesis in the standard conditions sufficient for effective reverse transcription of rabbit globin mRNA and of poly(A) in the presence of oligo(dT) primers. After a mild alkaline treatment of the mRNA and subsequent polyadenylation of the 3'-termini of the generated fragments with ATP:RNA adenyltrans-ferase from E.coli the poly(A)(+) polyribonucleotides are able to serve as templates for reverse transcription in the presence of oligo(dT) and the reverse transcriptase. A conclusion is made that a "structural stop" exists in mitochondrial mRNA non-translable regions adjacent to the poly(A) terminal sequence. The "structural stop" is suggested to be caused by post-transcriptional modification of mRNA (methylation, etc.) or by a particularly stable secondary structure in this region of the mRNA molecules.


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