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PMID: 10606665 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro evidence for the interaction of tRNA(3)(Lys) with U3 during the first strand transfer of HIV-1 reverse transcription.

Nucleic acids research ·Vol. 28 ·No. 2 ·2000-01-15 ·Pages 634-40

Brulé F, Bec G, Keith G, Le Grice SF, Roques BP, Ehresmann B, Ehresmann C, Marquet R

Abstract

Over the course of its evolution, HIV-1 has taken maximum advantage of its tRNA(3)(Lys)primer by utilizing it in several steps of reverse transcription. Here, we have identified a conserved nonanucleotide sequence in the U3 region of HIV-1 RNA that is complementary to the anticodon stem of tRNA(3)(Lys). In order to test its possible role in the first strand transfer reaction, we applied an assay using a donor RNA corresponding to the 5'-part and an acceptor RNA spanning the 3'-part of HIV-1 RNA. In addition, we constructed two acceptor RNAs in which the nonanucleotide sequence complementary to tRNA(3)(Lys)was either substituted (S) or deleted (Delta). We used either natural tRNA(3)(Lys)or an 18 nt DNA as primer and measured the efficiency of (-) strand strong stop DNA transfer in the presence of wild-type, S or Delta acceptor RNA. Mutations in U3 did not decrease the transfer efficiency when reverse transcription was primed with the 18mer DNA. However, they significantly reduced the strand transfer efficiency in the tRNA(3)(Lys)-primed reactions. This reduction was also observed in the presence of nucleocapsid protein. These results suggest that tRNA(3)(Lys)increases (-) strand strong stop transfer by interacting with the U3 region of the genomic RNA. Sequence comparisons suggest that such long range interactions also exist in other lentiviruses.

MeSH Terms
Base Sequence HIV Reverse Transcriptase/metabolism Nucleic Acid Conformation Polymerase Chain Reaction RNA, Transfer, Lys/metabolism RNA, Viral/chemistry,metabolism Transcription, Genetic
Chemicals
RNA, Transfer, Lys RNA, Viral HIV Reverse Transcriptase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Brulé F
Unité Propre de Recherche No. 9002 du CNRS, Institut de Biologie Moléculaire et Cellulaire, 15 rue René Descartes, 67084 Strasbourg Cedex, France,
Bec G
Keith G
Le Grice S F
Roques B P
Ehresmann B
Ehresmann C
Marquet R
References (38)
38 references, click to expand
  1. Divalent cation modulation of the ribonuclease functions of human immunodeficiency virus reverse transcriptase.
    Biochemistry. 1995 Aug 8;34(31):9936-43 PMID: 7543283
  2. RNA-dependent DNA polymerase in virions of RNA tumour viruses.
    Nature. 1970 Jun 27;226(5252):1209-11 PMID: 4316300
  3. Specific initiation and switch to elongation of human immunodeficiency virus type 1 reverse transcription require the post-transcriptional modifications of primer tRNA3Lys.
    EMBO J. 1996 Feb 15;15(4):917-24 PMID: 8631312
  4. Molecular analysis of the second template switch during reverse transcription of the HIV RNA template.
    Biochemistry. 1996 Aug 13;35(32):10549-57 PMID: 8756711
  5. Human immunodeficiency virus Type 1 nucleocapsid protein (NCp7) directs specific initiation of minus-strand DNA synthesis primed by human tRNA(Lys3) in vitro: studies of viral RNA molecules mutated in regions that flank the primer binding site.
    J Virol. 1996 Aug;70(8):4996-5004 PMID: 8764006
  6. Primer selection by HIV-1 reverse transcriptase on RNA-tRNA(3Lys) and DNA-tRNA(3Lys) hybrids.
    J Mol Biol. 1996 Aug 23;261(3):315-21 PMID: 8780773
  7. Initiation of (-) strand DNA synthesis from tRNA(3Lys) on lentiviral RNAs: implications of specific HIV-1 RNA-tRNA(3Lys) interactions inhibiting primer utilization by retroviral reverse transcriptases.
    Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10063-8 PMID: 8816751
  8. Human immunodeficiency virus type 1 nucleocapsid protein promotes efficient strand transfer and specific viral DNA synthesis by inhibiting TAR-dependent self-priming from minus-strand strong-stop DNA.
    J Virol. 1997 Jul;71(7):5178-88 PMID: 9188585
  9. Posttranscriptional modification of retroviral primers is required for late stages of DNA replication.
    Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7210-5 PMID: 9207070
  10. Binding of RNA template to a complex of HIV-1 reverse transcriptase/primer/template.
    Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11279-84 PMID: 9326600
  11. Primer tRNAs for reverse transcription.
    J Virol. 1997 Nov;71(11):8087-95 PMID: 9343157
  12. Mutational analysis of the tRNA3Lys/HIV-1 RNA (primer/template) complex.
    Nucleic Acids Res. 1998 Mar 1;26(5):1198-204 PMID: 9469827
  13. Genetic analysis of the U5-PBS of a novel HIV-1 reveals multiple interactions between the tRNA and RNA genome required for initiation of reverse transcription.
    RNA. 1998 Apr;4(4):394-406 PMID: 9630246
  14. The yeast Ty3 retrotransposon contains a 5'-3' bipartite primer-binding site and encodes nucleocapsid protein NCp9 functionally homologous to HIV-1 NCp7.
    EMBO J. 1998 Aug 17;17(16):4873-80 PMID: 9707446
  15. Nucleic-acid-chaperone activity of retroviral nucleocapsid proteins: significance for viral replication.
    Trends Biochem Sci. 1998 Aug;23(8):297-301 PMID: 9757830
  16. The native structure of the human immunodeficiency virus type 1 RNA genome is required for the first strand transfer of reverse transcription.
    Virology. 1998 Sep 30;249(2):211-8 PMID: 9791013
  17. Role of post-transcriptional modifications of primer tRNALys,3 in the fidelity and efficacy of plus strand DNA transfer during HIV-1 reverse transcription.
    J Biol Chem. 1999 Feb 12;274(7):4412-20 PMID: 9933645
  18. Structural basis for the specificity of the initiation of HIV-1 reverse transcription.
    EMBO J. 1999 Feb 15;18(4):1038-48 PMID: 10022845
  19. Modified nucleotides of tRNA(3Lys) modulate primer/template loop-loop interaction in the initiation complex of HIV-1 reverse transcription.
    J Biol Chem. 1993 Dec 5;268(34):25269-72 PMID: 7503978
  20. Transactivation of the minus-strand DNA transfer by nucleocapsid protein during reverse transcription of the retroviral genome.
    EMBO J. 1994 Feb 15;13(4):973-81 PMID: 7509280
  21. RNA-dependent DNA polymerase in virions of Rous sarcoma virus.
    Nature. 1970 Jun 27;226(5252):1211-3 PMID: 4316301
  22. A detailed model of reverse transcription and tests of crucial aspects.
    Cell. 1979 Sep;18(1):93-100 PMID: 509527
  23. Small finger protein of avian and murine retroviruses has nucleic acid annealing activity and positions the replication primer tRNA onto genomic RNA.
    EMBO J. 1988 Jun;7(6):1777-83 PMID: 2458920
  24. Rapid purification of homodimer and heterodimer HIV-1 reverse transcriptase by metal chelate affinity chromatography.
    Eur J Biochem. 1990 Jan 26;187(2):307-14 PMID: 1688798
  25. Dimerization of human immunodeficiency virus (type 1) RNA: stimulation by cations and possible mechanism.
    Nucleic Acids Res. 1991 May 11;19(9):2349-57 PMID: 1645868
  26. First large scale chemical synthesis of the 72 amino acid HIV-1 nucleocapsid protein NCp7 in an active form.
    Biochem Biophys Res Commun. 1991 Oct 31;180(2):1010-8 PMID: 1953705
  27. Viral RNA annealing activities of human immunodeficiency virus type 1 nucleocapsid protein require only peptide domains outside the zinc fingers.
    Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6472-6 PMID: 1631144
  28. Mechanism of DNA strand transfer reactions catalyzed by HIV-1 reverse transcriptase.
    Science. 1992 Nov 13;258(5085):1112-8 PMID: 1279806
  29. Two short basic sequences surrounding the zinc finger of nucleocapsid protein NCp10 of Moloney murine leukemia virus are critical for RNA annealing activity.
    Nucleic Acids Res. 1993 Feb 25;21(4):823-9 PMID: 8451185
  30. Interactions between HIV-1 nucleocapsid protein and viral DNA may have important functions in the viral life cycle.
    Nucleic Acids Res. 1993 Feb 25;21(4):831-9 PMID: 8383840
  31. Comparison of deoxyoligonucleotide and tRNA(Lys-3) as primers in an endogenous human immunodeficiency virus-1 in vitro reverse transcription/template-switching reaction.
    J Biol Chem. 1994 May 20;269(20):14672-80 PMID: 7514178
  32. Trans-activation of the 5' to 3' viral DNA strand transfer by nucleocapsid protein during reverse transcription of HIV1 RNA.
    C R Acad Sci III. 1993 Aug;316(8):763-71 PMID: 7519118
  33. Recombinant HIV-1 nucleocapsid protein accelerates HIV-1 reverse transcriptase catalyzed DNA strand transfer reactions and modulates RNase H activity.
    Biochemistry. 1994 Nov 22;33(46):13817-23 PMID: 7524664
  34. Human immunodeficiency virus nucleocapsid protein accelerates strand transfer of the terminally redundant sequences involved in reverse transcription.
    J Biol Chem. 1994 Dec 16;269(50):31491-5 PMID: 7989315
  35. Truncating alpha-helix E' of p66 human immunodeficiency virus reverse transcriptase modulates RNase H function and impairs DNA strand transfer.
    J Biol Chem. 1995 Mar 31;270(13):7068-76 PMID: 7535765
  36. Initiation of reverse transcription of HIV-1: secondary structure of the HIV-1 RNA/tRNA(3Lys) (template/primer).
    J Mol Biol. 1995 Mar 24;247(2):236-50 PMID: 7707372
  37. tRNAs as primer of reverse transcriptases.
    Biochimie. 1995;77(1-2):113-24 PMID: 7541250
  38. Construction of a type 1 human immunodeficiency virus that maintains a primer binding site complementary to tRNA(His).
    J Virol. 1996 Feb;70(2):966-75 PMID: 8551637
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/28.2.634
Published
2000-01-15
Pages
634-40
Language
English
Region
England
NLM ID
0411011
PMCID
PMC102502
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/28.2.634 PMC Full Text View journal details
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