HIV-1 reverse transcriptase discriminates against non-self tRNA primers

被引:43
作者
Essink, BBO [1 ]
Das, AT [1 ]
Berkhout, B [1 ]
机构
[1] UNIV AMSTERDAM,ACAD MED CTR,DEPT HUMAN RETROVIROL,NL-1105 AZ AMSTERDAM,NETHERLANDS
关键词
reverse transcription; HIV-1; retrovirus; tRNA primer; primer-binding site;
D O I
10.1006/jmbi.1996.0638
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (PBSLys3) is replaced by sequences complementary to tRNA(lie), 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. (C) 1996 Academic Press Limited
引用
收藏
页码:243 / 254
页数:12
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