| Aptamer redesigned tRNA is nonfunctional and degraded in cells. | |
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MedLine Citation:
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PMID: 14681579 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An RNA aptamer derived from tRNA(Gln) isolated in vitro and a rationally redesigned tRNA(Gln) were used to address the relationship between structure and function of tRNA(Gln) aminoacylation in Escherichia coli. Two mutant tRNA(Gln) sequences were studied: an aptamer that binds 26-fold tighter to glutaminyl-tRNA synthetase than wild-type tRNA(Gln) in vitro, redesigned in the variable loop, and a mutant with near-normal aminoacylation kinetics for glutamine, redesigned to contain a long variable arm. Both mutants were tested in a tRNA(Gln) knockout strain of E. coli, but neither supported knockout cell growth. It was later found that both mutant tRNAs were present in very low amounts in the cell. These results reveal the difference between in vitro and in vivo studies, demonstrating the complexities of in vivo systems that have not been replicated in vitro. |
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Authors:
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Dennis Lee; William H McClain |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: RNA (New York, N.Y.) Volume: 10 ISSN: 1355-8382 ISO Abbreviation: RNA Publication Date: 2004 Jan |
Date Detail:
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Created Date: 2003-12-18 Completed Date: 2004-01-29 Revised Date: 2013-06-09 |
Medline Journal Info:
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Nlm Unique ID: 9509184 Medline TA: RNA Country: United States |
Other Details:
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Languages: eng Pagination: 7-11 Citation Subset: IM |
Affiliation:
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Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706-1569, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acylation Amino Acyl-tRNA Synthetases / genetics, metabolism* Cells, Cultured Escherichia coli / enzymology*, genetics Glutamine / metabolism Kinetics Mutagenesis, Site-Directed Mutation Nucleic Acid Conformation Peptide Chain Initiation, Translational / genetics* Plasmids RNA, Bacterial / chemistry* RNA, Transfer, Gln / genetics*, metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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GM42123/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/RNA, Bacterial; 0/RNA, Transfer, Gln; 56-85-9/Glutamine; EC 6.1.1.-/Amino Acyl-tRNA Synthetases; EC 6.1.1.18/glutaminyl-tRNA synthetase |
| Comments/Corrections | |
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