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Initiation of in vitro reverse transcription from tRNA Lys3 on HIV‐1 or HIV‐2 RNAs by both type 1 and 2 reverse transcriptases
Author(s) -
Boulmé Florence,
Freund Frédéric,
Litvak Simon
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00649-8
Subject(s) - reverse transcriptase , complementary dna , biology , rna , microbiology and biotechnology , transfer rna , dna , elongation factor , elongation , rna directed dna polymerase , nucleotide , genetics , gene , ribosome , materials science , ultimate tensile strength , metallurgy
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)·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.