HIV-1 reverse transcriptase discriminates against non-self tRNA primers.
The interactions between the Reverse Transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) and the natural tRNA(Lys3) primer for initiation of viral DNA synthesis were examined. We constructed a set of HIV-1 RNA templates in which the wild-type primer binding site (PBS(Lys3)) is replaced by sequences complementary to tRNA(lle), tRNA(Lys1,2), tRNA(Phe), tRNA(Pro) or tRNA(Trp) and tested the ability of RT enzymes of different retroviral species to initiate cDNA synthesis from self versus non-self tRNA primers. We demonstrate that initiation of HIV-1 reverse transcription is a specific process that is most efficient with the self tRNA(Lys3) primer. Interestingly, the property of HIV-1 RT to discriminate against non-self tRNA primers is lost upon extension of the tRNA by only two deoxyribonucleotides. Furthermore, selective tRNA priming by HIV-1 RT was not observed with viral RNA-tRNA(Lys3) duplexes isolated from HIV-1 virion particles, suggesting that the majority of tRNA(Lys3) primers annealed to viral RNA in particles is extended by a variable number of deoxyribonucleotides. This result indicates that reverse transcription is initiated relatively early in nascently assembled virions.
Oude Essink
B B
BB
Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, The Netherlands.
Das
A T
AT
Berkhout
B
B
D003854:Deoxyribonucleotides; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D006801:Humans; D012313:RNA; D012343:RNA, Transfer; D012367:RNA, Viral; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014771:Virion
D003854:Deoxyribonucleotides; C034477:RNA primers; D012367:RNA, Viral; D012313:RNA; D012343:RNA, Transfer; D054303:HIV Reverse Transcriptase