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Science 28 July 2006:
Vol. 313. no. 5786, pp. 518 - 520
DOI: 10.1126/science.1127422

Reports

Transcript-Assisted Transcriptional Proofreading

Nikolay Zenkin,1* Yulia Yuzenkova,1 Konstantin Severinov1,2,3*

Fidelity of template-dependent nucleic acid synthesis is the main determinant of stable heredity and error-free gene expression. The mechanism (or mechanisms) ensuring fidelity of transcription by DNA-dependent RNA polymerases (RNAPs) is not fully understood. Here, we show that the 3' end–proximal nucleotide of the nascent transcript stimulates hydrolysis of the penultimate phosphodiester bond by providing active groups and coordination bonds to the RNAP active center. This stimulation is much higher in the case of misincorporated nucleotide. We show that during transcription elongation, the hydrolytic reaction stimulated by misincorporated nucleotides proofreads most of the misincorporation events and thus serves as an intrinsic mechanism of transcription fidelity.

1 Waksman Institute, Rutgers University, Piscataway, NJ 08854, USA.
2 Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ 08854, USA.
3 Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, 123182 Russia.

* To whom correspondence should be addressed at Waksman Institute, 190 Frelinghuysen Road, Piscataway, NJ 08854, USA. E-mail: nicserzen{at}mail.ru (N.Z.); severik{at}waksman.rutgers.edu (K.S.)

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THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Fluctuations, Pauses, and Backtracking in DNA Transcription.
M. Voliotis, N. Cohen, C. Molina-Paris, and T. B. Liverpool (2008)
Biophys. J. 94, 334-348
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Analysis of Promoter Targets for Escherichia coli Transcription Elongation Factor GreA In Vivo and In Vitro.
E. Stepanova, J. Lee, M. Ozerova, E. Semenova, K. Datsenko, B. L. Wanner, K. Severinov, and S. Borukhov (2007)
J. Bacteriol. 189, 8772-8785
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Nonrandom variations in human cancer ESTs indicate that mRNA heterogeneity increases during carcinogenesis.
M. Brulliard, D. Lorphelin, O. Collignon, W. Lorphelin, B. Thouvenot, E. Gothie, S. Jacquenet, V. Ogier, O. Roitel, J.-M. Monnez, et al. (2007)
PNAS 104, 7522-7527
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Science. ISSN 0036-8075 (print), 1095-9203 (online)