Human immunodeficiency virus uses tRNALys,3 as primer for reverse transcription in HeLa‐CD4+ cells

FEBS Letters

Koken, Sabine E.C.¹  van Wamel, Jeroen L.B.¹  Das, Atze T.¹  Oude Essink, Belinda B.¹  Berkhout, Ben¹ 

[1] Department of Virology, University of Amsterdam, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands

关键词: Reverse transcription;    tRNALys;    3;    Human immunodeficiency virus type 1;    RNA-RNA interaction;   

DOI  :  10.1016/0014-5793(94)80238-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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 and 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 [(1992) 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.

【 授权许可】

Unknown