On the fidelity of transcription by Escherichia coli ribonucleic acid polymerase

Springgate, Clark F.; Loeb, Lawrence A.

doi:10.1016/s0022-2836(75)80060-x

Cited by 76 publications

(45 citation statements)

References 24 publications

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“…RNA polymerases, however, have a much higher fidelity of incorporation than do DNA primases (43). Since each event of primer synthesis involves a competition between the opposing movements of the associated helicase translocation activity and the polymerization of ribonucleotides, it seems reasonable that DNA primases have evolved such that they sacrifice fidelity for speed of primer synthesis.…”

Section: Discussionmentioning

confidence: 99%

Template Recognition and Ribonucleotide Specificity of the DNA Primase of Bacteriophage T7

Kusakabe

Richardson

1997

Journal of Biological Chemistry

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The 63-kDa gene 4 DNA primase of phage T7 catalyzes the synthesis of oligoribonucleotides on single-stranded DNA templates. At the sequence, 5-GTC-3, the primase synthesizes the dinucleotide pppAC; the cytidine residue of the recognition sequence is cryptic. Only tetraribonucleotides function as primers, but the specificity for the third and fourth position is not as stringent with a preference of CMP > AMP > > UMP > GMP. The predominant recognition sites on M13 DNA are 5-(G/T)G-GTC-3 and 5-GTGTC-3. Synthesis is usually limited to tetranucleotides, but T7 primase can synthesize longer oligoribonucleotides on templates containing long stretches of guanosine residues 5 to the recognition sequence. The specificity beyond the first two positions of the primer increases as the length of the template on the 3-side of 5-GTC-3 increases. On an oligonucleotide having 20 3-flanking cytidine residues GMP is incorporated at the third position; incorporation is reduced 4-fold when the flanking sequence reaches 65 residues, and little is incorporated on M13 templates. The presence of the 56-kDa gene 4 helicase decreases the incorporation of GMP on long templates. We propose that pausing is required for the incorporation of less preferred nucleotides and that pausing is decreased by the ability of the primase to translocate 5 to 3 on templates having long 3-flanking sequences.

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Section: Discussionmentioning

confidence: 99%

Template Recognition and Ribonucleotide Specificity of the DNA Primase of Bacteriophage T7

Kusakabe

Richardson

1997

Journal of Biological Chemistry

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“…In vivo measurements of errors in gene expression in bacteria have been estimated to occur at a rate of 10 −4 to 10 −5 per nucleotide during transcription and ≈10 −3 to 10 −4 per codon during translation. These error rates are significantly higher than that attributed to the highfidelity replication process, where the frequency of errors is as low as 10 −8 to 10 −9 per nucleotide (8,9,(12)(13)(14)(15)(16)(17)(18)(19)(20).…”

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confidence: 83%

Visualizing high error levels during gene expression in living bacterial cells

Meyerovich

Mamou

Ben-Yehuda

2010

Proc. Natl. Acad. Sci. U.S.A.

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To monitor inaccuracy in gene expression in living cells, we designed an experimental system in the bacterium Bacillus subtilis whereby spontaneous errors can be visualized and quantified at a single-cell level. Our strategy was to introduce mutations into a chromosomally encoded gfp allele, such that errors in protein production are reported in real time by the formation of fluorescent GFP molecules. The data reveal that the amount of errors can greatly exceed previous estimates, and that the error rate increases dramatically at lower temperatures and during stationary phase. Furthermore, we demonstrate that when facing an antibiotic threat, an increase in error level is sufficient to allow survival of bacteria carrying a mutated antibiotic-resistance gene. We propose that bacterial gene expression is error prone, frequently yielding protein molecules that differ slightly from the sequence specified by their DNA, thus generating a cellular reservoir of nonidentical protein molecules. This variation may be a key factor in increasing bacterial fitness, expanding the capability of an isogenic population to face environmental challenges.Bacillus subtilis | translational errors | variations in living cells | translation fidelity D NA is duplicated with remarkable fidelity to ensure that accurate genetic information is transmitted from one generation to the next. This information is passed from DNA to RNA and from RNA to protein during gene expression; however, the accuracy of these downstream events is relatively less understood. Although RNA and proteins are generally considered short-lived noninherited molecules, several diseases are now known to arise from errors occurring during transcription and translation (1-3), implying that faithful transfer of genetic information from DNA to proteins is crucial for maintaining proper cellular functions.

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“…Cases (a) and (b) are known and have been described in a number of studies (8)(9)(10)(11)(12)(13)(14). In this communication we wish to propose a mechanism that could accomplish the function (c).…”

Section: Introductionmentioning

confidence: 99%

A possible mechanism responsible for the correction of transcription errors

Volloch¹,

Rits²,

Tumerman³

1979

Nucl Acids Res

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Nucleoside triphosphate phosphohydrolase (NTPase) activity was found in a preparation of E. Coli RNA polymerase. This enzymatic activity is capaable of hydrolysing all four ribonucleoside triphosphates to the nucleoside diphosphates. However, during in vitro RNA synthesis directed by poly(dC) or poly(dT), only the non-complementary nucleoside triphosphate of the same heterocyclic class was hydrolysed. No incorporation of the non-complementary precursor into RNA could be detected in these experiments. When another RNA polymerase preparation, devoid of NTPase activity, was employed, there was no hydrolysis of any nucleoside triphosphate and significant incorporation of non-complemtary precursor into RNA was observed.These observations lead us to the conclusion that NTPase, acting in conjunction with RNA polymerase, has the function of correcting errors in transcription.

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On the fidelity of transcription by Escherichia coli ribonucleic acid polymerase

Cited by 76 publications

References 24 publications

Template Recognition and Ribonucleotide Specificity of the DNA Primase of Bacteriophage T7

Template Recognition and Ribonucleotide Specificity of the DNA Primase of Bacteriophage T7

Visualizing high error levels during gene expression in living bacterial cells

A possible mechanism responsible for the correction of transcription errors

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