Yusupova G, Lanchy J M, Yusupov M, Keith G, Le Grice S F, Ehresmann C, Ehresmann B, Marquet R
Unité Propre de Recherche no. 9002 du Centre National de la Recherche Scientifique Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
J Mol Biol. 1996 Aug 23;261(3):315-21. doi: 10.1006/jmbi.1996.0463.
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-tRNA3Lys, and DNA-tRNA3Lys complexes by HIV-1 RT. When the 3' end of tRNA3Lys is annealed to oligoribonucleotides, tRNA3Lys, but not the complementary RNAs, is extended by HIV-1 RT, indicating that tRNA3Lys is efficiently used as primer and RNA as template. An opposite primer usage is observed when tRNA3Lys is annealed to complementary oligodeoxyribonucleotides. In this case, the oligodeoxyribonucleotides are efficiently used as primer and tRNA3Lys as template. This result indicates that the nature of nucleic acid bound to tRNA3Lys determines which strand of the RNA-tRNA3Lys and DNA-tRNA3Lys hybrids is extended by HIV-1 RT. When an oligoribonucleotide is annealed to an unmodified transcript of tRNA3Lys, both nucleic acids are extended by HIV-1 RT, indicating that specific selection of tRNA3Lys as primer requires the post-transcriptional modifications of tRNA3Lys.
在人类免疫缺陷病毒1型（HIV-1）的基因组RNA逆转录为双链DNA的过程中，逆转录酶（RT）必须以RNA-RNA、DNA-RNA、RNA-DNA和DNA-DNA杂交体作为引物模板。在本研究中，我们检测了HIV-1 RT对RNA-tRNA3Lys和DNA-tRNA3Lys复合物的延伸情况。当tRNA3Lys的3'末端与寡核糖核苷酸退火时，HIV-1 RT可延伸tRNA3Lys，而非互补RNA，这表明tRNA3Lys可有效用作引物，RNA用作模板。当tRNA3Lys与互补寡脱氧核糖核苷酸退火时，观察到相反的引物使用情况。在这种情况下，寡脱氧核糖核苷酸可有效用作引物，tRNA3Lys用作模板。这一结果表明，与tRNA3Lys结合的核酸的性质决定了HIV-1 RT延伸RNA-tRNA3Lys和DNA-tRNA3Lys杂交体的哪一条链。当寡核糖核苷酸与未修饰的tRNA3Lys转录本退火时，两种核酸均可被HIV-1 RT延伸，这表明tRNA3Lys作为引物的特异性选择需要tRNA3Lys的转录后修饰。
