TY - JOUR
T1 - Primer selection by HIV-1 reverse transcriptase on RNA-tRNA3(Lys) and DNA-tRNA3(Lys) hybrids
AU - Yusupova, Gulnara
AU - Lanchy, Jean Marc
AU - Yusupov, Marat
AU - Keith, Gérard
AU - Le Grice, Stuart F.J.
AU - Ehresmann, Chantal
AU - Ehresmann, Bernard
AU - Marquet, Roland
N1 - Funding Information:
Philippe Dumas, Catherine Isel and Eugene Skripkin are thanked for fruitful discussions. The tRNA transcript was a gift from C. Isel. This work was supported by the French ‘‘Agence Nationale de Recherches sur le SIDA’’ (ANRS). G.Y. was supported by a fellowship from CNRS in the frame of the Russian–French scientific cooperation, and M.Y. was supported by a grant from Genset (Paris). S. Le G. is supported by grant AI31147 from the National Institutes of Health.
PY - 1996/8/23
Y1 - 1996/8/23
N2 - 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).
AB - 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).
KW - HIV-1
KW - Polymerase
KW - Retrovirus
KW - Reverse transcriptase
KW - Transfer RNA
UR - http://www.scopus.com/inward/record.url?scp=0030598921&partnerID=8YFLogxK
U2 - 10.1006/jmbi.1996.0463
DO - 10.1006/jmbi.1996.0463
M3 - Editorial
C2 - 8780773
AN - SCOPUS:0030598921
SN - 0022-2836
VL - 261
SP - 315
EP - 321
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 3
ER -
