M3i

@article{,

title = {In vitro evidence for the interaction of tRNA(3)(Lys) with U3 during the first strand transfer of HIV-1 reverse transcription},

author = {F Brule and G Bec and G Keith and S F Le Grice and B P Roques and B Ehresmann and C Ehresmann and R Marquet},

url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10606665},

isbn = {10606665},

year  = {2000},

date = {2000-01-01},

journal = {Nucleic Acids Res},

volume = {28},

number = {2},

pages = {634-640},

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.},

note = {1362-4962 

Journal Article},

keywords = {Base Sequence  HIV-1 Reverse Transcriptase/*metabolism  Nucleic Acid Conformation  Polymerase Chain Reaction  RNA, Genetic, Lys/*metabolism  RNA, MARQUET, Non-U.S. Gov't  *Transcription, Transfer, Unité ARN, Viral/chemistry/*metabolism  Support},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Specific initiation and switch to elongation of human immunodeficiency virus type 1 reverse transcription require the post-transcriptional modifications of primer tRNA3Lys},

author = {C Isel and J M Lanchy and S F Le Grice and C Ehresmann and B Ehresmann and R Marquet},

url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8631312},

isbn = {8631312},

year  = {1996},

date = {1996-01-01},

journal = {EMBO J},

volume = {15},

number = {4},

pages = {917-924},

abstract = {Initiation of RNA-dependent DNA synthesis by retroviral reverse transcriptases is generally considered as unspecific. In the case of human immunodeficiency virus type 1 (HIV-1), the natural primer is tRNA3Lys. We recently found evidence of complex interactions between tRNA3Lys and HIV-1 RNA that may be involved in the priming process. In this study, we compare the ability of natural and unmodified synthetic tRNA3Lys and 18mer oligoribo- and oligodeoxyribonucleotides complementary to the viral primer binding site to initiate replication of HIV-1 RNA using either homologous or heterologous reverse transcriptases. We show that HIV-1 RNA, HIV-1 reverse transcriptase and primer tRNA3Lys form a specific initiation complex that differs from the unspecific elongation complex formed when an oligodeoxyribonucleotide is used as primer. Modified nucleosides of tRNA3Lys are required for efficient initiation and transition to elongation. Transition from initiation to elongation, but not initiation of reverse transcription itself, is facilitated by extended primer-template interactions. Elongation, but not initiation of reverse transcription, is inhibited by Mn2+, which further differentiates these two different functional states of reverse transcriptase. These results define initiation of reverse transcription as a target to block viral replication.},

note = {0261-4189 

Journal Article},

keywords = {Cell-Free System  Gene Expression Regulation, Genetic  *Virus Replication, Lys/*metabolism  RNA, MARQUET, Non-U.S. Gov't  Templates, Transfer, Unité ARN, Viral  HIV-1/*genetics  HIV-1 Reverse Transcriptase  RNA, Viral/metabolism  RNA-Directed DNA Polymerase/*metabolism  Support},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Modified nucleotides of tRNA(3Lys) modulate primer/template loop-loop interaction in the initiation complex of HIV-1 reverse transcription},

author = {C Isel and R Marquet and G Keith and C Ehresmann and B Ehresmann},

url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7503978},

isbn = {7503978},

year  = {1993},

date = {1993-01-01},

journal = {J Biol Chem},

volume = {268},

number = {34},

pages = {25269-25272},

abstract = {In all retroviruses, reverse transcription is primed by a tRNA whose 3' end 18 nucleotides are complementary to the so called viral primer binding site. Previous work showed that reverse transcription of HIV-1 RNA is initiated by tRNA(3Lys). Using a variety of chemical and enzymatic structural probes, we investigated the interactions between HIV-1 RNA and its natural primer tRNA(3Lys). In addition to the predictable contacts between the viral primer binding site and the 3' end of tRNA(3Lys), a specific interaction takes place between an A-rich loop located upstream of the primer binding site region and the anticodon loop of tRNA(3Lys). This AAAA/Umcm5s2UUU loop-loop interaction is not observed when the natural primer is replaced by an in vitro synthesized tRNA(3Lys) transcript. Furthermore, dethiolation of the modified nucleotide mcm5s2U at position 34 of tRNA(3Lys) strongly destabilizes this interaction. Sequence and structure comparisons indicate that the primer/template loop-loop interaction is conserved in all HIV-1 isolates, and possibly also in HIV-2 and SIV.},

note = {0021-9258 

Journal Article},

keywords = {Anticodon/genetics/metabolism  Base Sequence  Binding Sites  Comparative Study  DNA Primers  HIV-1/genetics/*metabolism  HIV-1 Reverse Transcriptase  HIV-2/genetics/metabolism  Molecular Sequence Data  Nucleic Acid Conformation  RNA, Genetic, Genetic  *Transcription, Lys/*metabolism  RNA, MARQUET, Non-U.S. Gov't  Templates, Transfer, Unité ARN, Viral/*biosynthesis  RNA-Directed DNA Polymerase/*metabolism  SIV/genetics/metabolism  Support},

pubstate = {published},

tppubtype = {article}

}