Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes

@article{Kozak1986PointMD,
  title={Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes},
  author={Marilyn Sue Kozak},
  journal={Cell},
  year={1986},
  volume={44},
  pages={283-292}
}
  • M. Kozak
  • Published 1986
  • Biology, Medicine
  • Cell
By analyzing the effects of single base substitutions around the ATG initiator codon in a cloned preproinsulin gene, I have identified ACCATGG as the optimal sequence for initiation by eukaryotic ribosomes. [...] Key Result Mutations within that sequence modulate the yield of proinsulin over a 20-fold range.Expand
At least six nucleotides preceding the AUG initiator codon enhance translation in mammalian cells.
  • M. Kozak
  • Biology, Medicine
  • Journal of molecular biology
  • 1987
TLDR
Analysis of a new set of mutants now reveals that sequences slightly farther upstream are also influential, the optimal context for initiation being (GCC)GCCA/GCC a/gCCAUGG, the repeating GCC motif. Expand
Modulation of translational efficiency by contextual nucleotides flanking a baculovirus initiator AUG codon.
TLDR
Experimental results suggest an optimal initiator context of 5'-N N N N n N N A U G A a/c/g N-3' for efficient translation initiation in lepidopteran cells. Expand
tRNAi(met) functions in directing the scanning ribosome to the start site of translation.
TLDR
An anticodon: codon interaction between tRNAi(met) as part of the scanning ribosome and the first AUG must function in directing the ribosomes to the eukaryotic initiator region. Expand
AUG context and mRNA translation in vaccinia virus
TLDR
The efficiency of translation was not affected significantly by nucleotide substitutions in positions –3 and +4, and despite the high occurrence of purine nucleotides in those positions in vaccinia mRNAs, no effect of those positions was noted in conditions where â-Gal was overexpressed. Expand
Sequence and structural features associated with translational initiator regions in yeast--a review.
TLDR
Compared the translational initiator regions of 131 yeast genes appears to indicate that basic features associated with yeast leader regions are consistent with a general mechanism of initiation of protein synthesis in eukaryotes, as proposed by the ribosomal 'scanning' model, but perhaps only basic featuresassociated with ribosome recognition of an AUG start codon are intact. Expand
Evidence for involvement of trans-acting factors in selection of the AUG start codon during eukaryotic translational initiation
TLDR
Findings indicate that trans-acting factors may play a role in AUG start site selection during translational initiation by eukaryotic ribosomes. Expand
Selection of the 5'-proximal translation initiation site is influenced by mRNA and eIF-2 concentrations.
A cDNA clone of the influenza virus NS (non-structural protein) gene in a vector carrying a bacteriophage T7 RNA polymerase promoter was manipulated so as to reiterate the initiation site to give twoExpand
Molecular Mechanism of Scanning and Start Codon Selection in Eukaryotes
  • A. Hinnebusch
  • Biology, Medicine
  • Microbiology and Molecular Reviews
  • 2011
TLDR
A robust molecular model now exists, describing the roles of initiation factors, notably eukaryotic initiation factor 1 and eIF1A, in stabilizing an “open” conformation of the 40S subunit with Met-tRNAiMet bound in a low-affinity state conducive to scanning and in triggering rearrangement into a “closed’ conformation incompatible with scanning. Expand
Effect of mutations downstream of the internal ribosome entry site on initiation of poliovirus protein synthesis
Initiation of poliovirus translation is mediated by a large, structured segment of the 5' nontranslated region known as the internal ribosome entry site (IRES) and normally occurs 155 nucleotidesExpand
The immediate downstream codon strongly influences the efficiency of utilization of eukaryotic translation initiation codons.
TLDR
With AUG as the initiation codon, the pattern of response to mutations in the +4 and +5 positions was qualitatively similar, but the quantitative effects were smaller, thus the optimum downstream context for initiation is A/CUGGAU. Expand
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References

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Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes.
  • M. Kozak
  • Biology, Medicine
  • Nucleic acids research
  • 1981
TLDR
The observed asymmetry suggests that purines in positions -3 and +4 might facilitate recognition of the AUG condon during formation of initiation complexes and a modified version of the scanning mechanism is proposed. Expand
Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.
  • M. Kozak
  • Biology, Medicine
  • Nucleic acids research
  • 1984
TLDR
The sequence CCAGCCAUG (G) thus emerges as a consensus sequence for eukaryotic initiation sites, and the extent to which the ribosome binding site in a given mRNA matches the -1 to -5 consensus sequence varies. Expand
Influences of mRNA secondary structure on initiation by eukaryotic ribosomes.
  • M. Kozak
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1986
Oligonucleotides designed to create hairpin structures were inserted upstream from the ATG initiator codon in several plasmids that encode preproinsulin, and the effects on translation were monitoredExpand
Initiation of translation at internal AUG codons in mammalian cells
TLDR
The results raise the possibility that mammalian ribosomes can reinitiate translation at an AUG codon after previously initiating, and terminating, at an upstream site. Expand
Selection of initiation sites by eucaryotic ribosomes: effect of inserting AUG triplets upstream from the coding sequence for preproinsulin.
  • M. Kozak
  • Biology, Medicine
  • Nucleic acids research
  • 1984
TLDR
Recombinant plasmids that direct synthesis of rat preproinsulin under the direction of the SV40 early promoter have been used to probe the mechanism of initiation of translation, finding that ribosomes initiate at the first AUG, terminate, and then reinitiate at the AUG that directly precedes the pre Proinsulin coding sequence. Expand
Point mutations close to the AUG initiator codon affect the efficiency of translation of rat preproinsulin in vivo
To determine whether sequence context influences the ability of an AUG triplet to be recognized as an initiator codon by eukaryotic ribosomes, single nucleotide changes were introduced near theExpand
Inability of circular mRNA to attach to eukaryotic ribosomes
TLDR
The experiments described below fulfill the simple prediction that circularisation of a message should abolish its ability to interact with ribosomes, and the threading mechanism is consistent with the mono-cistronic character of eukaryotic messages. Expand
Initiation of translation of the cauliflower mosaic virus genome from a polycistronic mRNA: evidence from deletion mutagenesis
TLDR
Results suggest translation from a polycistronic mRNA by a ‘relay race’ mechanism whereby ribosomes after passing a termination codon re‐initiate protein synthesis at the nearest AUG codon. Expand
Translation of insulin-related polypeptides from messenger RNAs with tandemly reiterated copies of the ribosome binding site
TLDR
Plasmids have been constructed containing reiterated copies of a 66 bp fragment that includes the AUG initiator codon of preproinsulin, and the position of an AUG triplet relative to the 5' end of the mRNA appears to be important in identifying it as a functional initiators codon. Expand
Mutation of a termination codon affects src initiation.
TLDR
It is suggested that the wild-type src message produces two polypeptides, a very small (nine-amino acid) peptide that is initiated at the gag AUG and the 60,000-dalton src protein that is initiate at the src AUG. Expand
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