Publications
2003
Frugier M, Giege R, Schimmel P
RNA recognition by designed peptide fusion creates Journal Article
In: Proc Natl Acad Sci U S A, vol. 100, no. 13, pp. 7471-7475, 2003, ISBN: 12796515, (0027-8424 Journal Article).
Abstract | Links | BibTeX | Tags: Amino Acid Motifs Amino Acid Sequence Amino Acyl-tRNA Ligases/*chemistry Base Sequence Binding Sites Catalytic Domain Escherichia coli/genetics/metabolism *Genetic Techniques Kinetics Models, FRUGIER, Genetic Molecular Sequence Data Mutation Nucleic Acid Conformation Peptides/*chemistry Protein Binding Protein Structure, Non-U.S. Gov't Support, P.H.S. Time Factors, Tertiary RNA/chemistry Recombinant Fusion Proteins/metabolism Support, U.S. Gov't, Unité ARN
@article{,
title = {RNA recognition by designed peptide fusion creates },
author = {M Frugier and R Giege and P Schimmel},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12796515},
isbn = {12796515},
year = {2003},
date = {2003-01-01},
journal = {Proc Natl Acad Sci U S A},
volume = {100},
number = {13},
pages = {7471-7475},
abstract = {The genetic code was established through aminoacylations of RNA substrates that emerged as tRNAs. The 20 aminoacyl-tRNA synthetases (one for each amino acid) are ancient proteins, the active-site domain of which catalyzes formation of an aminoacyl adenylate that subsequently reacts with the 3' end of bound tRNA. Binding of tRNA depends on idiosyncratic (to the particular synthetase) domains and motifs that are fused to or inserted into the conserved active-site domain. Here we take the domain for synthesis of alanyl adenylate and fuse it to "artificial" peptide sequences (28 aa) that were shown previously to bind to the acceptor arm of tRNAAla. Certain fusions confer aminoacylation activity on tRNAAla and on hairpin microhelices modeled after its acceptor stem. Aminoacylation was sensitive to the presence of a specific G:U base pair known to be a major determinant of tRNAAla identity. Aminoacylation efficiency and specificity also depended on the specific peptide sequence. The results demonstrate that barriers to RNA-specific aminoacylations are low and can be achieved by relatively simple peptide fusions. They also suggest a paradigm for rationally designed specific aminoacylations based on peptide fusions.},
note = {0027-8424
Journal Article},
keywords = {Amino Acid Motifs Amino Acid Sequence Amino Acyl-tRNA Ligases/*chemistry Base Sequence Binding Sites Catalytic Domain Escherichia coli/genetics/metabolism *Genetic Techniques Kinetics Models, FRUGIER, Genetic Molecular Sequence Data Mutation Nucleic Acid Conformation Peptides/*chemistry Protein Binding Protein Structure, Non-U.S. Gov't Support, P.H.S. Time Factors, Tertiary RNA/chemistry Recombinant Fusion Proteins/metabolism Support, U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2001
Wilhelm M, Uzun O, Mules E H, Gabriel A, Wilhelm F X
Polypurine tract formation by Ty1 RNase H Journal Article
In: J Biol Chem, vol. 276, no. 50, pp. 47695-47701, 2001, ISBN: 11595735, (0021-9258 Journal Article).
Abstract | Links | BibTeX | Tags: Base Sequence Binding Sites DNA/metabolism DNA Primers/pharmacology Hydrolysis Molecular Sequence Data Mutation Protein Binding *Purines RNA/metabolism RNA-Directed DNA Polymerase/*chemistry/*metabolism Recombinant Proteins/metabolism *Retroelements Ribonuclease H, Calf Thymus/*chemistry/*genetics/metabolism Support, Non-U.S. Gov't Support, P.H.S. Time Factors, U.S. Gov't, Unité ARN
@article{,
title = {Polypurine tract formation by Ty1 RNase H},
author = {M Wilhelm and O Uzun and E H Mules and A Gabriel and F X Wilhelm},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11595735},
isbn = {11595735},
year = {2001},
date = {2001-01-01},
journal = {J Biol Chem},
volume = {276},
number = {50},
pages = {47695-47701},
abstract = {To better understand the mechanism by which Ty1 RNase H creates the polypurine tract (PPT) primer, we have demonstrated the polymerase-dependent hydrolytic activity of Ty1 reverse transcriptase (RT) during minus-strand synthesis. Using RNase H and polymerase mutants of the recombinant Ty1 RT protein, we show that the two domains of Ty1 RT can act independently of one another. Our results indicate that RNA/DNA substrates containing a short RNA PPT, which serve as primers for plus-strand DNA synthesis, are relatively resistant to RNase H cleavage. RNA substrates with a correct 5' end but with 3' end extending beyond the plus-strand initiation site were cleaved specifically to generate the correct 3' end of the PPT. Using long RNA/DNA duplexes containing the PPT, we show that Ty1 RT is able to make specific internal cleavages that could generate the plus-strand primer with correct 5' and 3' ends. Long RNA/DNA duplexes with mutations in the PPT or in a U-rich region upstream of the PPT, which abolish plus-strand initiation in vivo, were not cleaved specifically at the 5' end of the PPT. Our work demonstrates that the in vitro enzyme can recapitulate key processes that control proper replication in vivo.},
note = {0021-9258
Journal Article},
keywords = {Base Sequence Binding Sites DNA/metabolism DNA Primers/pharmacology Hydrolysis Molecular Sequence Data Mutation Protein Binding *Purines RNA/metabolism RNA-Directed DNA Polymerase/*chemistry/*metabolism Recombinant Proteins/metabolism *Retroelements Ribonuclease H, Calf Thymus/*chemistry/*genetics/metabolism Support, Non-U.S. Gov't Support, P.H.S. Time Factors, U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}