Initiation of (-) strand DNA synthesis from tRNALys 3 on lentiviral RNAs: Implications of specific HIV-1 RNA-tRNALys 3 interactions inhibiting primer utilization by retroviral reverse transcriptases

Eric J. Arts, Scott R. Stetor, Xuguang Li, Jason W. Rausch, Kathyrn J. Howard, Bernard Ehresmann, Thomas W. North, Birgitta M. Wöhrl, Roger S. Goody, Mark A. Wainberg, Stuart F J Le Grice

Research output: Contribution to journalArticle

68 Scopus citations

Abstract

Initiation of minus (-) strand DNA synthesis was examined on templates containing R, U5, and primer-binding site regions of the human immunodeficiency virus type 1 (HIV-1), feline immunodeficiency virus (FIV), and equine infectious anemia virus (EIAV) genomic RNA. DNA synthesis was initiated from (i) an oligoribonucleotide complementary to the primer-binding sites, (ii) synthetic tRNALys 3, and (iii) natural tRNALys 3, by the reverse transcriptases of HIV-1, FIV, EIAV, simian immunodeficiency virus, HIV type 2 (HIV-2), Moloney murine leukemia virus, and avian myeloblastosis virus. All enzymes used an oligonucleotide on wild-type HIV-1 RNA, whereas only a limited number initiated (-) strand DNA synthesis from either tRNALys 3. In contrast, all enzymes supported efficient tRNALys 3-primed (-) strand DNA synthesis on the genomes of FIV and EIAV. This may be in part attributable to the observation that the U5-inverted repeat stem-loop of the EIAV and FIV genomes lacks an A-rich loop shown with HIV-1 to interact with the U-rich tRNA anticodon loop. Deletion of this loop in HIV-1 RNA, or disrupting a critical loop-loop complex by tRNALys 3 extended by 9 nt, restored synthesis of HIV-1 (-) strand DNA from primer tRNALys 3 by all enzymes. Thus, divergent evolution of lentiviruses may have resulted in different mechanisms to use the same host tRNA for initiation of reverse transcription.

Original languageEnglish (US)
Pages (from-to)10063-10068
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number19
StatePublished - Sep 17 1996
Externally publishedYes

Keywords

  • HIV-1 reverse transcription
  • Retroviral replication

ASJC Scopus subject areas

  • General
  • Genetics

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    Arts, E. J., Stetor, S. R., Li, X., Rausch, J. W., Howard, K. J., Ehresmann, B., North, T. W., Wöhrl, B. M., Goody, R. S., Wainberg, M. A., & Le Grice, S. F. J. (1996). Initiation of (-) strand DNA synthesis from tRNALys 3 on lentiviral RNAs: Implications of specific HIV-1 RNA-tRNALys 3 interactions inhibiting primer utilization by retroviral reverse transcriptases. Proceedings of the National Academy of Sciences of the United States of America, 93(19), 10063-10068.