Two genes specifying model mRNAs of minimal size and coding capacity, with or without the Shine-Dalgarno (SD) sequence, were assembled, cloned, and transcribed in high yields. These mRNAs, as well as synthetic polynucleotides, phage MS2 RNA, and a deoxyoctanucleotide complementary to the 3' end of 16S rRNA were used to study the mechanism of translation initiation in vitro. Escherichia coli 30S ribosomal subunits interact with all these nucleic acids, albeit with different affinities; the affinity for the mRNA with the SD sequence (Ka approximately 2 x 10(7) M-1) is more than an order of magnitude higher than that for the mRNA lacking this sequence. The initiation factors are equally required, regardless of the presence of the SD sequence, for 30S and 70S initiation complex formation and for mRNA translation, but the initiation factors do not affect the SD interaction or the binding of the mRNAs to the ribosomes. The SD interaction is also mechanistically irrelevant for 30S initiation complex formation and is not essential for translation in vitro or for the selection of the mRNA reading frame. It is suggested that the function of the SD interaction is to ensure a high concentration of the initiation triplet near the ribosomal ...Continue Reading
Initial rate kinetic analysis of the mechanism of initiation complex formation and the role of initiation factor IF-3
A single base change in the Shine-Dalgarno region of 16S rRNA of Escherichia coli affects translation of many proteins
Specialized ribosome system: preferential translation of a single mRNA species by a subpopulation of mutated ribosomes in Escherichia coli
Chemical synthesis and in vivo hyperexpression of a modular gene coding for Escherichia coli translational initiation factor IF1
The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites
Effect of Escherichia coli initiation factors on the kinetics of N-Acphe-tRNAPhe binding to 30S ribosomal subunits. A fluorescence stopped-flow study
The ribosome binding sites recognized by E. coli ribosomes have regions with signal character in both the leader and protein coding segments
An insight into the possible mechanism of working of two-cistronic gene expression systems and rational designing of newer systems
30S ribosomal subunits with fragmented 16S RNA: a new approach for structure and function study of ribosomes
Ribosome-mRNA contact sites at different stages of translation initiation as revealed by cross-linking of model mRNAs
Inducible high expression of the Escherichia coli infC gene subcloned behind a bacteriophage T7 promoter
Initiation of Escherichia coli ribosomes on matrix coupled mRNAs studied by optical biosensor technique
Translation during cold adaptation does not involve mRNA-rRNA base pairing through the downstream box
Potential for interdependent development of tRNA determinants for aminoacylation and ribosome decoding
Secondary structure of the ribosome binding site determines translational efficiency: a quantitative analysis
Translation of mRNAs with degenerate initiation triplet AUU displays high initiation factor 2 dependence and is subject to initiation factor 3 repression
The "allosteric three-site model" of elongation cannot be confirmed in a well-defined ribosome system from Escherichia coli
Truncated elongation factor G lacking the G domain promotes translocation of the 3' end but not of the anticodon domain of peptidyl-tRNA
Release factor RF3 in E.coli accelerates the dissociation of release factors RF1 and RF2 from the ribosome in a GTP-dependent manner
Fast recycling of Escherichia coli ribosomes requires both ribosome recycling factor (RRF) and release factor RF3
Selective stimulation of translation of leaderless mRNA by initiation factor 2: evolutionary implications for translation
Site-directed cross-linking of mRNA analogues to the Escherichia coli ribosome; identification of 30S ribosomal components that can be cross-linked to the mRNA at various points 5' with respect to the decoding site
Lack of a 5' non-coding region in Tn1721 encoded tetR mRNA is associated with a low efficiency of translation and a short half-life in Escherichia coli
Interaction of mRNA with the Escherichia coli ribosome: accessibility of phosphorothioate-containing mRNA bound to ribosomes for iodine cleavage
The mRNA for the 23S rRNA methylase encoded by the ermE gene of Saccharopolyspora erythraea is translated in the absence of a conventional ribosome-binding site
Alternative occupancy of a dual ribosomal binding site by mRNA affected by translation initiation factors
The highly efficient translation initiation region from the Escherichia coli rpsA gene lacks a shine-dalgarno element
Depletion of free 30S ribosomal subunits in Escherichia coli by expression of RNA containing Shine-Dalgarno-like sequences
Haloferax volcanii, a prokaryotic species that does not use the Shine Dalgarno mechanism for translation initiation at 5'-UTRs
Impact of P-Site tRNA and antibiotics on ribosome mediated protein folding: studies using the Escherichia coli ribosome
Engineered Expression Vectors Significantly Enhanced the Production of 2-Keto-D-gluconic Acid by Gluconobacter oxidans
Characterization of a lipoprotein, NilC, required by Xenorhabdus nematophila for mutualism with its nematode host
Colicin E3 cleavage of 16S rRNA impairs decoding and accelerates tRNA translocation on Escherichia coli ribosomes
Detection of Escherichia coli ribosome binding at translation initiation sites in the absence of tRNA
A target-unrelated peptide in an M13 phage display library traced to an advantageous mutation in the gene II ribosome-binding site
Inhibition of translation initiation complex formation by GE81112 unravels a 16S rRNA structural switch involved in P-site decoding
Accurate translocation of mRNA by the ribosome requires a peptidyl group or its analog on the tRNA moving into the 30S P site
Large-scale movement of elongation factor G and extensive conformational change of the ribosome during translocation
Selective translation of leaderless mRNAs by specialized ribosomes generated by MazF in Escherichia coli
Initiation factors IF1 and IF2 synergistically remove peptidyl-tRNAs with short polypeptides from the P-site of translating Escherichia coli ribosomes
Analysis of SD sequences in completed microbial genomes: non-SD-led genes are as common as SD-led genes
Escherichia coli initiation factor 3 protein binding to 30S ribosomal subunits alters the accessibility of nucleotides within the conserved central region of 16S rRNA
The anti-Shine-Dalgarno region in Escherichia coli 16S ribosomal RNA is not essential for the correct selection of translational starts
Fluorescence study of the topology of messenger RNA bound to the 30S ribosomal subunit of Escherichia coli
Nature of the ribosomal mRNA track: analysis of ribosome-binding sites containing different sequences and secondary structures
Streptomycin interferes with conformational coupling between codon recognition and GTPase activation on the ribosome
Reinitiation of protein synthesis in Escherichia coli can be induced by mRNA cis-elements unrelated to canonical translation initiation signals
The effect of ribosomal protein S1 from Escherichia coli and Micrococcus luteus on protein synthesis in vitro by E. coli and Bacillus subtilis
Directed evolution of ribosomal protein S1 for enhanced translational efficiency of high GC Rhodopseudomonas palustris DNA in Escherichia coli
S1 ribosomal protein functions in translation initiation and ribonuclease RegB activation are mediated by similar RNA-protein interactions: an NMR and SAXS analysis
Structure of a 30S pre-initiation complex stalled by GE81112 reveals structural parallels in bacterial and eukaryotic protein synthesis initiation pathways
Polyamine stimulation of the synthesis of oligopeptide-binding protein (OppA). Involvement of a structural change of the Shine-Dalgarno sequence and the initiation codon aug in oppa mRNA
Translational repression of the Escherichia coli alpha operon mRNA: importance of an mRNA conformational switch and a ternary entrapment complex
Unique localization of the plastid-specific ribosomal proteins in the chloroplast ribosome small subunit provides mechanistic insights into the chloroplastic translation
The pentatricopeptide repeat protein PGR3 is required for the translation of petL and ndhG by binding their 5'UTRs
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