|
|
|
|||||||
|
|||||||||
J Biol Chem, Vol. 275, Issue 16, 12306-12312, April 21, 2000
From the Initiation of human immunodeficiency virus-1
(HIV-1) reverse transcription requires formation of a complex
containing the viral RNA (vRNA),
tRNA3Lys and reverse
transcriptase (RT). The vRNA and the primer
tRNA3Lys form several
intermolecular interactions in addition to annealing of the primer 3'
end to the primer binding site (PBS). These interactions are crucial
for the efficiency and the specificity of the initiation of reverse
transcription. However, as they are located upstream of the PBS, they
must unwind as DNA synthesis proceeds. Here, the dynamics of the
complex during initiation of reverse transcription was followed by
enzymatic probing. Our data revealed reciprocal effects of the tertiary
structure of the vRNA·tRNA3Lys
complex and reverse transcriptase (RT) at a distance from the polymerization site. The structure of the initiation complex allowed RT
to interact with the template strand up to 20 nucleotides upstream from
the polymerization site. Conversely, nucleotide addition by RT modified
the tertiary structure of the complex at 10-14 nucleotides from the
catalytic site. The viral sequences became exposed at the surface of
the complex as they dissociated from the tRNA following primer
extension. However, the counterpart tRNA sequences became buried inside
the complex. Surprisingly, they became exposed when mutations prevented
the intermolecular interactions in the initial complex, indicating that
the fate of the tRNA depended on the tertiary structure of the initial complex.
Dynamics of the HIV-1 Reverse Transcription Complex during
Initiation of DNA Synthesis*
§,,,,, and
UPR 9002 du CNRS, IBMC, 67084 Strasbourg
cedex, France and ¶ Division of Infectious Diseases, Case
Western Reserve University School of Medicine,
Cleveland, Ohio 44106
*
This work was supported by the Agence Nationale de Recherche
sur le SIDA (ANRS) and by a Biomed II grant from the European Union.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: UPR 9002 du CNRS,
IBMC, 15 rue René Descartes, 67084 Strasbourg cedex, France. Tel.: 33 3 88 41 70 91; Fax: 33 3 88 60 22 18; E-mail:
marquet@ibmc.u- strasbg.fr.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 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]
|
|
|
|
 |
|
 C. Dash, T. S. Fisher, V. R. Prasad, and S. F. J. Le Grice Examining Interactions of HIV-1 Reverse Transcriptase with Single-stranded Template Nucleotides by Nucleoside Analog Interference J. Biol. Chem., September 22, 2006; 281(38): 27873 - 27881. [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]
|
|
|
|
|
|
Y. Iwatani, A. E. Rosen, J. Guo, K. Musier-Forsyth, and J. G. Levin Efficient Initiation of HIV-1 Reverse Transcription in Vitro. REQUIREMENT FOR RNA SEQUENCES DOWNSTREAM OF THE PRIMER BINDING SITE ABROGATED BY NUCLEOCAPSID PROTEIN-DEPENDENT PRIMER-TEMPLATE INTERACTIONS J. Biol. Chem., April 11, 2003; 278(16): 14185 - 14195. [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]
|
|
|
|
|
|
J. G. Julias, M. J. McWilliams, S. G. Sarafianos, E. Arnold, and S. H. Hughes Mutations in the RNase H domain of HIV-1 reverse transcriptase affect the initiation of DNA synthesis and the specificity of RNase H cleavage in vivo PNAS, July 9, 2002; 99(14): 9515 - 9520. [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]
|
|
|
|
|
|
J. T. Miller, B. Ehresmann, U. Hubscher, and S. F. J. Le Grice A Novel Interaction of tRNALys,3 with the Feline Immunodeficiency Virus RNA Genome Governs Initiation of Minus Strand DNA Synthesis J. Biol. Chem., July 13, 2001; 276(29): 27721 - 27730. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lavigne, L. Polomack, and H. Buc Structures of Complexes Formed by HIV-1 Reverse Transcriptase at a Termination Site of DNA Synthesis J. Biol. Chem., August 10, 2001; 276(33): 31439 - 31448. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |