Initiation of in vitro reverse transcription from tRNA(Lys3) on HIV-1 or HIV-2 RNAs by both type 1 and 2 reverse transcriptases.
HIV reverse transcription is initiated from a cellular tRNA partially associated with the retroviral genome. Here we studied homologous HIV-2 cDNA synthesis using natural or synthetic primers. With natural tRNA(Lys3), synthesis of early products comprising nucleotides +5 to +7 preceded the elongation step leading to synthesis of (-) strong-stop cDNA. In the presence of a poly(A) x oligo(dT) trap, no full-length product was observed while early products were still present, showing a transition between initiation and elongation. With DNA primers only an unspecific elongation was found. Our data show a similar mechanism of reverse transcription initiation by HIV-1 and HIV-2 reverse transcriptases. Furthermore, using a heterologous system we found that HIV-1 RNA, in contrast to data reported in the literature, was an excellent template for HIV-2 reverse transcriptase.
EP-630, CNRS-Université Victor Ségalen Bordeaux 2, IFR 66 Pathologies Infectieuses, France.
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Boulmé
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Freund
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Litvak
D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics*; D015498:HIV-2 / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D012313:RNA; D012346:RNA, Transfer, Amino Acyl / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; C034477:RNA primers; D012346:RNA, Transfer, Amino Acyl; D012367:RNA, Viral; D012313:RNA; C072416:reverse transcriptase, Human immunodeficiency virus 2; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase