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

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Abstract

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 tRNALys,3 molecule as primer. This is based on sequence complementarity between the 3' end of tRNALys,3 and the primer-binding site (PBS) on HIV-1 genomic RNA. Recent biochemical analyses indicated that tRNALys,3 is indeed incorporated into viral particles. Interestingly, tRNALys,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 tRNALys,3 as primer. Most importantly, we obtained revertants of one such PBS-mutant, which had restored a wild-type PBS sequence. This tRNALys,3-mediated repair demonstrates a general requirement for this primer in HIV-1 reverse transcription. © 1994.

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Das, A. T., Koken, S. E. C., Oude Essink, B. B., van Wamel, J. L. B., & Berkhout, B. (1994). Human immunodeficiency virus uses tRNALys,3 as primer for reverse transcription in HeLa-CD4+ cells. FEBS Letters, 341(1), 49–53. https://doi.org/10.1016/0014-5793(94)80238-6

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