HUMAN-IMMUNODEFICIENCY-VIRUS USES TRNA(LYS,3) AS PRIMER FOR REVERSE TRANSCRIPTION IN HELA-CD4(+) CELLS

被引:12
作者
DAS, AT [1 ]
KOKEN, SEC [1 ]
ESSINK, BBO [1 ]
VANWAMEL, JLB [1 ]
BERKHOUT, B [1 ]
机构
[1] UNIV AMSTERDAM,ACAD MED CTR,DEPT VIROL,1105 AZ AMSTERDAM,NETHERLANDS
关键词
REVERSE TRANSCRIPTION; TRNA(LYS; 3); HUMAN IMMUNODEFICIENCY VIRUS TYPE 1; RNA-RNA INTERACTION;
D O I
10.1016/0014-5793(94)80238-6
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
Significant amounts of different tRNA molecules are present in retroviral particles, but one specific tRNA species functions as primer in reverse transcription. It is generally believed that the HIV-1 virus uses the tRNA(Lys,3) molecule as primer. This is based on sequence complementarity between the 3' end of tRNA(Lys,3) the primer-binding site (PBS) on HIV-1 genomic RNA. Recent biochemical analyses indicated that tRNA(Lys,3) is indeed incorporated into viral particles. Interestingly, tRNA(Lys,3) could not be detected in virions produced by HeLa-CD4(+) cells [Biochem. Biophys. Res. Commun. 185, 1105-1115]. In order to test whether alternative tRNA molecules can function as primer in HIV replication, we performed a series of experiments based on the observation that tRNA primer sequences are inherited by the viral progeny. We cultured HIV-1 for prolonged periods of time in HeLa-CD4(+) cells, but did not detect sequence changes in the PBS region. Furthermore, we found PBS-mutants to be replication-incompetent, again suggesting that HIV-1 solely uses tRNA(Lys,3) as primer. Most importantly, we obtained revertants of one such PBS-mutant, which had restored a wild-type PBS sequence. This tRNA(Lys,3)-mediated repair demonstrates a general requirement for this primer in HIV-1 reverse transcription.
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收藏
页码:49 / 53
页数:5
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