Evidence for a unique first position codon-anticodon mismatch in vivo

Toth, Matthew J.; Murgola, Emanuel J.; Schimmel, Paul

doi:10.1016/0022-2836(88)90152-0

Cited by 54 publications

(53 citation statements)

References 15 publications

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“…For example, a rpsD mutation increased the third position G d G misreading of the UGG tryptophan codon by tRNA Cys GCA during translation of ribosomal protein S6 (Bouadloun et al 1983) and increased the misincorporation of cysteine into the T7 0.3 protein as well (Rice et al 1984). A first position U d G misreading of GGC (Gly) by tRNA Ser GCU was also significantly increased by the rpsD mutation (Toth et al 1988). It has, however, also been reported that the rpsD mutation had no effect on the first position A d C misreading of CGU (Arg) by tRNA Cys GCA (Bouadloun et al 1983) or on the misincorporation of histidine into the Qb protein (Khazaie et al 1984).…”

Section: An Rpsd Mutation Increases Misreading Of Several Near-cognatmentioning

confidence: 84%

“…The misincorporation frequency is approximately the ratio of the enzyme activity expressed from the mutant to that of the wild-type control. Toth et al (1988) estimated the frequency that tRNA Ser GCU misreads a GGC glycine codon as about 10 À3 per codon using a mutant form of b-lactamase.…”

Section: Introductionmentioning

confidence: 99%

“…A third approach uses a reporter gene bearing a mutation altering an essential amino acid (Toth et al 1988;Cornut and Willson 1991). Misincorporation of the wild-type amino acid at the mutant codon would restore enzymatic activity.…”

Section: Introductionmentioning

confidence: 99%

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The frequency of translational misreading errors in E. coli is largely determined by tRNA competition

Kramer¹,

Farabaugh²

2006

RNA

421

516

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Estimates of missense error rates (misreading) during protein synthesis vary from 10 À3 to 10 À4 per codon. The experiments reporting these rates have measured several distinct errors using several methods and reporter systems. Variation in reported rates may reflect real differences in rates among the errors tested or in sensitivity of the reporter systems. To develop a more accurate understanding of the range of error rates, we developed a system to quantify the frequency of every possible misreading error at a defined codon in Escherichia coli. This system uses an essential lysine in the active site of firefly luciferase. Mutations in Lys529 result in up to a 1600-fold reduction in activity, but the phenotype varies with amino acid. We hypothesized that residual activity of some of the mutant genes might result from misreading of the mutant codons by tRNA Lys UUU , the cognate tRNA for the lysine codons, AAA and AAG. Our data validate this hypothesis and reveal details about relative missense error rates of near-cognate codons. The error rates in E. coli do, in fact, vary widely. One source of variation is the effect of competition by cognate tRNAs for the mutant codons; higher error frequencies result from lower competition from low-abundance tRNAs. We also used the system to study the effect of ribosomal protein mutations known to affect error rates and the effect of error-inducing antibiotics, finding that they affect misreading on only a subset of near-cognate codons and that their effect may be less general than previously thought.

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Section: An Rpsd Mutation Increases Misreading Of Several Near-cognatmentioning

confidence: 84%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The frequency of translational misreading errors in E. coli is largely determined by tRNA competition

Kramer¹,

Farabaugh²

2006

RNA

421

516

View full text Add to dashboard Cite

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“…The E. coli IT41 parent strain is E. coli W3110 (14), a nonsuppressing strain, and therefore the behavior of E. coli IT41 at 42ЊC cannot be explained by the presence of a Ts amber suppressor phenotype. Cases of mistranslation of protein coding regions in mutagenesis experiments have been noted before (32). In this case, a serine (AGC) codon of a ␤-lactamase was changed to glycine codons GGC and GGA in order to produce a catalytically inactive protein.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning and expression of the spsB gene encoding an essential type I signal peptidase from Staphylococcus aureus

1996

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The gene, spsB, encoding a type I signal peptidase has been cloned from the gram-positive eubacterium Staphylococcus aureus. The gene encodes a protein of 191 amino acid residues with a calculated molecular mass of 21,692 Da. Comparison of the protein sequence with those of known type I signal peptidases indicates conservation of amino acid residues known to be important or essential for catalytic activity. The enzyme has been expressed to high levels in Escherichia coli and has been demonstrated to possess enzymatic activity against E. coli preproteins in vivo. Experiments whereby the spsB gene was transferred to a plasmid that is temperature sensitive for replication indicate that spsB is an essential gene. We identified an open reading frame immediately upstream of the spsB gene which encodes a type I signal peptidase homolog of 174 amino acid residues with a calculated molecular mass of 20,146 Da that is predicted to be devoid of catalytic activity.

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“…4). U36•G1 errors have been demonstrated for misreading of the Gly GGC codon by tRNA Ser GCU (Toth et al 1988). Given our observation of large differences in error frequencies among nearcognate tRNAs and the lack of data on the structure formed by other near-cognates, it is premature to conclude that other base mismatches can be constrained to interact in WatsonCrick geometry.…”

Section: Minimum Errors During Translation Is Surprisingly Lowmentioning

confidence: 99%

Studies of translational misreading in vivo show that the ribosome very efficiently discriminates against most potential errors

Manickam

Nag

Abbasi

et al. 2013

RNA

126

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Evidence for a unique first position codon-anticodon mismatch in vivo

Cited by 54 publications

References 15 publications

The frequency of translational misreading errors in E. coli is largely determined by tRNA competition

The frequency of translational misreading errors in E. coli is largely determined by tRNA competition

Molecular cloning and expression of the spsB gene encoding an essential type I signal peptidase from Staphylococcus aureus

Studies of translational misreading in vivo show that the ribosome very efficiently discriminates against most potential errors

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