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Mutations induced by bacteriophage T7 RNA polymerase and their effects on the composition of the T7 genome.
J Mol Biol 2000; 300(5):1057-65JM

Abstract

We show here that transcription by the bacteriophage T7 RNA polymerase increases the deamination of cytosine bases in the non-transcribed strand to uracil, causing C to T mutations in that strand. Under optimal conditions, the mutation frequency increases about fivefold over background, and is similar to that seen with the Escherichia coli RNA polymerase. Further, we found that a mutant T7 RNA polymerase with a slower rate of elongation caused more cytosine deaminations than its wild-type parent. These results suggest that promoting cytosine deamination in the non-transcribed strand is a general property of transcription in E. coli and is dependent on the length of time the transcription bubble stays open during elongation. To see if transcription-induced mutations have influenced the evolution of bacteriophage T7, we analyzed its genome for a bias in base composition. Our analysis showed a significant excess of thymine over cytosine bases in the highly transcribed regions of the genome. Moreover, the average value of this bias correlated well with the levels of transcription of different genomic regions. Our results indicate that transcription-induced mutations have altered the composition of bacteriophage T7 genome and suggest that this may be a significant force in genome evolution.

Authors+Show Affiliations

Department of Chemistry, Wayne State University, Detroit, MI 48202, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10903854

Citation

Beletskii, A, et al. "Mutations Induced By Bacteriophage T7 RNA Polymerase and Their Effects On the Composition of the T7 Genome." Journal of Molecular Biology, vol. 300, no. 5, 2000, pp. 1057-65.
Beletskii A, Grigoriev A, Joyce S, et al. Mutations induced by bacteriophage T7 RNA polymerase and their effects on the composition of the T7 genome. J Mol Biol. 2000;300(5):1057-65.
Beletskii, A., Grigoriev, A., Joyce, S., & Bhagwat, A. S. (2000). Mutations induced by bacteriophage T7 RNA polymerase and their effects on the composition of the T7 genome. Journal of Molecular Biology, 300(5), pp. 1057-65.
Beletskii A, et al. Mutations Induced By Bacteriophage T7 RNA Polymerase and Their Effects On the Composition of the T7 Genome. J Mol Biol. 2000 Jul 28;300(5):1057-65. PubMed PMID: 10903854.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mutations induced by bacteriophage T7 RNA polymerase and their effects on the composition of the T7 genome. AU - Beletskii,A, AU - Grigoriev,A, AU - Joyce,S, AU - Bhagwat,A S, PY - 2000/7/25/pubmed PY - 2000/9/2/medline PY - 2000/7/25/entrez SP - 1057 EP - 65 JF - Journal of molecular biology JO - J. Mol. Biol. VL - 300 IS - 5 N2 - We show here that transcription by the bacteriophage T7 RNA polymerase increases the deamination of cytosine bases in the non-transcribed strand to uracil, causing C to T mutations in that strand. Under optimal conditions, the mutation frequency increases about fivefold over background, and is similar to that seen with the Escherichia coli RNA polymerase. Further, we found that a mutant T7 RNA polymerase with a slower rate of elongation caused more cytosine deaminations than its wild-type parent. These results suggest that promoting cytosine deamination in the non-transcribed strand is a general property of transcription in E. coli and is dependent on the length of time the transcription bubble stays open during elongation. To see if transcription-induced mutations have influenced the evolution of bacteriophage T7, we analyzed its genome for a bias in base composition. Our analysis showed a significant excess of thymine over cytosine bases in the highly transcribed regions of the genome. Moreover, the average value of this bias correlated well with the levels of transcription of different genomic regions. Our results indicate that transcription-induced mutations have altered the composition of bacteriophage T7 genome and suggest that this may be a significant force in genome evolution. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/10903854/Mutations_induced_by_bacteriophage_T7_RNA_polymerase_and_their_effects_on_the_composition_of_the_T7_genome_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(00)93944-5 DB - PRIME DP - Unbound Medicine ER -