Enzymatic synthesis of DNA complementary to mitochondrial mRNA via reverse transcription.
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 myeloblastosis 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 adenyltransferase 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.
D000818:Animals; D004272:DNA, Mitochondrial / Q000096:biosynthesis*; D004277:DNA, Single-Stranded / Q000096:biosynthesis; D008930:Mitochondria, Liver / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D051381:Rats; D013698:Templates, Genetic
D004272:DNA, Mitochondrial; D004277:DNA, Single-Stranded; D012333:RNA, Messenger; D011061:Poly A; D011062:Polynucleotide Adenylyltransferase; D012194:RNA-Directed DNA Polymerase