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Folding interactions of an mRNA 5′‐UTR that regulates gene expression: Interaction with a regulatory tRNA
Author(s) -
Nelson Audrey Rose,
Henkin Tina,
Agris Paul F.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a537-b
Subject(s) - transfer rna , untranslated region , messenger rna , biology , gene , stem loop , five prime untranslated region , genetics , translational regulation , specifier , gene expression , microbiology and biotechnology , translation (biology) , rna , linguistics , philosophy , noun , noun phrase
Genes for aminoacyl‐tRNA synthetases (aaRS) in pathogenic gram positive bacteria (Streptococcus, Staphylococcus, Bacillus) are uniquely regulated through unacylated tRNA’s interaction with the 5′‐untranslated region (5′‐UTR) of the mRNA. The anticodon of the tRNA interacts with a codon sequence occurring in the ’Specifier Loop’ of the 5′‐UTR. Those aaRS genes, regulated through mRNA Specifier Loops, have evolved with codon sequences for different tRNAs in order to selectively control expression of the different aaRS genes. For the 5′‐UTR to positively affect gene expression through regulation of transcription, the mRNA folding interactions must be significantly stable to present the codon sequence for a productive binding to tRNA’s anticodon. A model, biomolecular system was engineered in which the interaction between two half molecules (Common and Specifier) would reconstitute the 5′‐UTR of the GlyRS mRNA, and as such could be analyzed for its structural and functional properties. Gel mobility shift analysis and fluorescence spectroscopy yielded experimental K d s for complex formation between Common and Specifier half molecules that demonstrated strong binding (10.5 ± 0.68 μM). The reconstituted 5′‐UTR of the GlyRS mRNA bound the anticodon stem and loop of tRNA Gly (ASL Gly GCC ) specifically and with a significant affinity (20.2 ± 1.38 μM). Thus, the biomolecular 5′‐UTR and ASL Gly GCC model mimics the interactive properties of transcriptional regulation deduced from molecular genetic experiments in vivo . NSF MCB‐9986011 (REU) and NIH GM23037 to P.F.A.