Brule F, Marquet R, Rong L, Wainberg M A, Roques B P, Grice S F Le, Ehresmann B, Ehresmann C
In: RNA, vol. 8, no. 1, pp. 8-15, 2002, ISBN: 11873759, (1355-8382 Journal Article).
Abstract | Links | BibTeX | Tags: Genetic, Genetic Transcription, Lys/*chemistry/genetics/metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/*chemistry/genetics/metabolism Support
@article{,
title = {Structural and functional properties of the HIV-1 RNA-tRNA(Lys)3 primer complex annealed by the nucleocapsid protein: comparison with the heat-annealed complex},
author = {F Brule and R Marquet and L Rong and M A Wainberg and B P Roques and S F Le Grice and B Ehresmann and C Ehresmann},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11873759},
isbn = {11873759},
year = {2002},
date = {2002-01-01},
journal = {RNA},
volume = {8},
number = {1},
pages = {8-15},
abstract = {The conversion of the single-stranded RNA genome into double-stranded DNA by virus-coded reverse transcriptase (RT) is an essential step of the retrovirus life cycle. In human immunodeficiency virus type 1 (HIV-1), RT uses the cellular tRNA(Lys)3 to initiate the (-) strand DNA synthesis. Placement of the primer tRNA(Lys)3 involves binding of its 3'-terminal 18 nt to a complementary region of genomic RNA termed PBS. However, the PBS sequence is not the unique determinant of primer usage and additional contacts are important. This placement is believed to be achieved in vivo by the nucleocapsid domain of Gag or by the mature protein NCp. Up to now, structural information essentially arose from heat-annealed primer-template complexes (Isel et al., J Mol Biol, 1995, 247:236-250; Isel et al., EMBO J, 1999, 18:1038-1048). Here, we investigated the formation of the primer-template complex mediated by NCp and compared structural and functional properties of heat- and NCp-annealed complexes. We showed that both heat- and NCp-mediated procedures allow comparable high yields of annealing. Then, we investigated structural features of both kinds of complexes by enzymatic probing, and we compared their relative efficiency in (-) strong stop DNA synthesis. We did not find any significant differences between these complexes, suggesting that information derived from the heat-annealed complex can be transposed to the NCp-mediated complex and most likely to complexes formed in vivo.},
note = {1355-8382
Journal Article},
keywords = {Genetic, Genetic Transcription, Lys/*chemistry/genetics/metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/*chemistry/genetics/metabolism Support},
pubstate = {published},
tppubtype = {article}
}
Tisne C, Rigourd M, Marquet R, Ehresmann C, Dardel F
In: RNA, vol. 6, no. 10, pp. 1403-1412, 2000, ISBN: 11073216, (1355-8382 Journal Article).
Abstract | Links | BibTeX | Tags: Base Sequence DNA, Biomolecular *Nucleic Acid Conformation RNA/*chemistry/genetics/metabolism RNA, Genetic Transcription, Genetic/*genetics Virus Replication, Lys/*chemistry/genetics/metabolism Structure-Activity Relationship Support, MARQUET, Molecular Molecular Sequence Data Mutation/genetics Nuclear Magnetic Resonance, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/biosynthesis/genetics Escherichia coli/genetics *Genetic Engineering HIV-1/*genetics/physiology Human Iodine/metabolism Models
@article{,
title = {NMR and biochemical characterization of recombinant human tRNA(Lys)3 expressed in Escherichia coli: identification of posttranscriptional nucleotide modifications required for efficient initiation of HIV-1 reverse transcription},
author = {C Tisne and M Rigourd and R Marquet and C Ehresmann and F Dardel},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11073216},
isbn = {11073216},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {10},
pages = {1403-1412},
abstract = {Reverse transcription of HIV-1 viral RNA uses human tRNA(Lys)3 as a primer. Some of the modified nucleotides carried by this tRNA must play a key role in the initiation of this process, because unmodified tRNA produced in vitro is only marginally active as primer. To provide a better understanding of the contribution of base modifications in the initiation complex, we have designed a recombinant system that allows tRNA(Lys)3 expression in Escherichia coli. Because of their high level of overexpression, some modifications are incorporated at substoichiometric levels. We have purified the two major recombinant tRNA(Lys)3 subspecies, and their modified nucleotide contents have been characterized by a combination of NMR and biochemical techniques. Both species carry psis, Ds, T, t6A, and m7G. Differences are observed at position 34, within the anticodon. One fraction lacks the 5-methylaminomethyl group, whereas the other lacks the 2-thio group. Although the s2U34-containing recombinant tRNA is a less efficient primer, it presents most of the characteristics of the mammalian tRNA. On the other hand, the mnm5U34-containing tRNA has a strongly reduced activity. Our results demonstrate that the modifications that are absent in E. coli (m2G10, psi27, m5C48, m5C49, and m1A58) as well as the mnm5 group at position 34 are dispensable for initiation of reverse transcription. In contrast, the 2-thio group at position 34 seems to play an important part in this process.},
note = {1355-8382
Journal Article},
keywords = {Base Sequence DNA, Biomolecular *Nucleic Acid Conformation RNA/*chemistry/genetics/metabolism RNA, Genetic Transcription, Genetic/*genetics Virus Replication, Lys/*chemistry/genetics/metabolism Structure-Activity Relationship Support, MARQUET, Molecular Molecular Sequence Data Mutation/genetics Nuclear Magnetic Resonance, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/biosynthesis/genetics Escherichia coli/genetics *Genetic Engineering HIV-1/*genetics/physiology Human Iodine/metabolism Models},
pubstate = {published},
tppubtype = {article}
}
Frugier M, Florentz C, Hosseini M W, Lehn J M, Giege R
Synthetic polyamines stimulate in vitro transcription by T7 RNA polymerase Journal Article
In: Nucleic Acids Res, vol. 22, no. 14, pp. 2784-2790, 1994, ISBN: 8052534, (0305-1048 Journal Article).
Abstract | Links | BibTeX | Tags: Bacteriophage T7/enzymology Base Sequence Comparative Study DNA-Directed RNA Polymerases/drug effects/*metabolism Kinetics Molecular Sequence Data Molecular Structure Nucleic Acid Conformation Oligodeoxyribonucleotides Polyamines/chemistry/*pharmacology Promoter Regions (Genetics) RNA, ERIANI, FLORENTZ, FRUGIER, Genetic Transcription, Genetic/*drug effects, Non-U.S. Gov't Templates, Transfer, Unité ARN, Val/*biosynthesis/chemistry Structure-Activity Relationship Support
@article{,
title = {Synthetic polyamines stimulate in vitro transcription by T7 RNA polymerase},
author = {M Frugier and C Florentz and M W Hosseini and J M Lehn and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8052534},
isbn = {8052534},
year = {1994},
date = {1994-01-01},
journal = {Nucleic Acids Res},
volume = {22},
number = {14},
pages = {2784-2790},
abstract = {The influence of nine synthetic polyamines on in vitro transcription with T7 RNA polymerase has been studied. The compounds used were linear or macrocyclic tetra- and hexaamine, varying in their size, shape and number of protonated groups. Their effect was tested on different types of templates, all presenting the T7 RNA promoter in a double-stranded form followed by sequences encoding short transcripts (25 to 35-mers) either on single- or double-stranded synthetic oligodeoxyribonucleotides. All polyamines used stimulate transcription of both types of templates at levels dependent on their size, shape, protonation degree, and concentration. For each compound, an optimal concentration could be defined; above this concentration, transcription inhibition occurred. Highest stimulation (up to 12-fold) was obtained by the largest cyclic compound called [38]N6C10.},
note = {0305-1048
Journal Article},
keywords = {Bacteriophage T7/enzymology Base Sequence Comparative Study DNA-Directed RNA Polymerases/drug effects/*metabolism Kinetics Molecular Sequence Data Molecular Structure Nucleic Acid Conformation Oligodeoxyribonucleotides Polyamines/chemistry/*pharmacology Promoter Regions (Genetics) RNA, ERIANI, FLORENTZ, FRUGIER, Genetic Transcription, Genetic/*drug effects, Non-U.S. Gov't Templates, Transfer, Unité ARN, Val/*biosynthesis/chemistry Structure-Activity Relationship Support},
pubstate = {published},
tppubtype = {article}
}