Primer selection by HIV-1 reverse transcriptase on RNA-tRNA3(Lys) and DNA-tRNA3(Lys) hybrids

Gulnara Yusupova, Jean Marc Lanchy, Marat Yusupov, Gérard Keith, Stuart F.J. Le Grice, Chantal Ehresmann, Bernard Ehresmann, Roland Marquet

Research output: Contribution to journalEditorial

Abstract

During reverse transcription of the genomic RNA of human immunodeficiency virus type 1 (HIV-1) into double-stranded DNA, reverse transcriptase (RT) must accommodate RNA-RNA, DNA-RNA, RNA-DNA and DNA-DNA hybrids as primer-template. In this study, we examined extension of RNA-tRNA3(Lys) and DNA-tRNA3(Lys) complexes by HIV-1 RT. When the 3' end of tRNA3(Lys) is annealed to oligoribonucleotides, tRNA3(Lys), but not the complementary RNAs, is extended by HIV-1 RT, indicating that tRNA3(Lys) is efficiently used as primer and RNA as template. An opposite primer usage is observed when tRNA3(Lys) is annealed to complementary oligodeoxyribonucleotides. In this case, the oligodeoxyribonucleotides are efficiently used as primer and tRNA3(Lys) as template. This result indicates that the nature of nucleic acid bound to tRNA3(Lys) determines which strand of the RNA-tRNA3(Lys) and DNA-tRNA3(Lys) hybrids is extended by HIV-1 RT. When an oligoribonucleotide is annealed to an unmodified transcript of tRNA3(Lys), both nucleic acids are extended by HIV-1 RT, indicating that specific selection of tRNA3(Lys) as primer requires the post-transcriptional modifications of tRNA3(Lys).

Original languageEnglish
Pages (from-to)315-321
Number of pages7
JournalJournal of Molecular Biology
Volume261
Issue number3
DOIs
StatePublished - Aug 23 1996

Keywords

  • HIV-1
  • Polymerase
  • Retrovirus
  • Reverse transcriptase
  • Transfer RNA

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