Nucleic Acids Research, 2001, Vol. 29, No. 13 2757-2765
© 2001 Oxford University Press
Initiation of HIV-2 reverse transcription: a secondary structure model of the RNA–tRNALys3 duplex
Human immunodeficiency virus type 2 (HIV-2) reverse transcription is initiated from cellular tRNALys3 partially annealed to the RNA viral genome at the primer binding site (PBS). This annealing involves interactions between two highly structured RNA molecules. In contrast to HIV-1, in which the reverse transcription initiation complex has been thoroughly studied, there is still little information regarding a possible model to describe the secondary structure of the template–primer complex in HIV-2. To determine whether HIV-2 RNA sequences flanking the PBS may specifically interact with the natural primer tRNA, we performed site-directed mutagenesis and enzymatic footprinting. An RNA fragment corresponding to the HIV-2 U5 RNA domain and tRNALys3 were probed either in their free form or in the binary complex. Important reactivity changes to nucleases were obtained upon complex formation. In addition to the canonical contacts between the viral PBS and the 3' end acceptor stem of tRNALys3, we identified two additional interacting domains: (i) the U-rich region of the anticodon loop with the A-rich sequence of the internal loop within the U5-prePBS region; (ii) nucleotides 48–54 from the T
C domain of tRNALys3 and the 240–247 region of viral U5-RNA. In view of these experimental data and sequence comparison between different HIV-2 isolates, we propose a model for the secondary structure of the HIV-2 template–primer initiation complex.
* To whom correspondence should be addressed. Tel: +33 5 57 57 17 60; Fax: +33 5 57 57 17 66; Email: laura.litvak{at}reger.u-bordeaux2.fr Present addresses:Frédéric Freund, Département de Microbiologie et d’Immunologie, Université de Montréal, Montréal, Québec H3C 3J7, CanadaFlorence Boulmé, Institute of Medical Biochemistry, Department of Molecular Biology, Vienna Biocenter. Dr Bohrgasse 9, 1030 Vienna, Austria
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Ooms, D. Cupac, T. E. M. Abbink, H. Huthoff, and B. Berkhout The availability of the primer activation signal (PAS) affects the efficiency of HIV-1 reverse transcription initiation Nucleic Acids Res., March 12, 2007; 35(5): 1649 - 1659. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. E. M. Abbink, N. Beerens, and B. Berkhout Forced Selection of a Human Immunodeficiency Virus Type 1 Variant That Uses a Non-Self tRNA Primer for Reverse Transcription: Involvement of Viral RNA Sequences and the Reverse Transcriptase Enzyme J. Virol., October 1, 2004; 78(19): 10706 - 10714. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. T. Miller, A. Khvorova, S. A. Scaringe, and S. F. J. Le Grice Synthetic tRNALys,3 as the replication primer for the HIV-1HXB2 and HIV-1Mal genomes Nucleic Acids Res., September 1, 2004; 32(15): 4687 - 4695. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Goldschmidt, J.-C. Paillart, M. Rigourd, B. Ehresmann, A.-M. Aubertin, C. Ehresmann, and R. Marquet Structural Variability of the Initiation Complex of HIV-1 Reverse Transcription J. Biol. Chem., August 20, 2004; 279(34): 35923 - 35931. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. A. Voronin and V. K. Pathak Frequent Dual Initiation in Human Immunodeficiency Virus-Based Vectors Containing Two Primer-Binding Sites: a Quantitative In Vivo Assay for Function of Initiation Complexes J. Virol., May 15, 2004; 78(10): 5402 - 5413. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Rigourd, V. Goldschmidt, F. Brule, C. D. Morrow, B. Ehresmann, C. Ehresmann, and R. Marquet Structure-function relationships of the initiation complex of HIV-1 reverse transcription: the case of mutant viruses using tRNAHis as primer Nucleic Acids Res., October 1, 2003; 31(19): 5764 - 5775. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Huthoff, K. Bugala, J. Barciszewski, and B. Berkhout On the importance of the primer activation signal for initiation of tRNAlys3-primed reverse transcription of the HIV-1 RNA genome Nucleic Acids Res., September 1, 2003; 31(17): 5186 - 5194. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Goldschmidt, C. Ehresmann, B. Ehresmann, and R. Marquet Does the HIV-1 primer activation signal interact with tRNA3Lys during the initiation of reverse transcription? Nucleic Acids Res., February 1, 2003; 31(3): 850 - 859. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kvaratskhelia, J. T. Miller, S. R. Budihas, L. K. Pannell, and S. F. J. Le Grice Identification of specific HIV-1 reverse transcriptase contacts to the viral RNA:tRNA complex by mass spectrometry and a primary amine selective reagent PNAS, December 10, 2002; 99(25): 15988 - 15993. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Goldschmidt, M. Rigourd, C. Ehresmann, S. F. J. Le Grice, B. Ehresmann, and R. Marquet Direct and Indirect Contributions of RNA Secondary Structure Elements to the Initiation of HIV-1 Reverse Transcription J. Biol. Chem., November 1, 2002; 277(45): 43233 - 43242. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Morris, M. Johnson, E. Stavnezer, and J. Leis Replication of Avian Sarcoma Virus In Vivo Requires an Interaction between the Viral RNA and the T{psi}C Loop of the tRNATrp Primer J. Virol., June 27, 2002; 76(15): 7571 - 7577. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Beerens and B. Berkhout The tRNA Primer Activation Signal in the Human Immunodeficiency Virus Type 1 Genome Is Important for Initiation and Processive Elongation of Reverse Transcription J. Virol., March 1, 2002; 76(5): 2329 - 2339. [Abstract] [Full Text] [PDF]
|
|