Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro.

Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro. Research Abstract Details 

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Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro. Abstract Text:

Ekaterina Stepanova,Jookyung Lee,Maria Ozerova,Ekaterina Semenova,Kirill Datsenko,Barry L Wanner,Konstantin Severinov,Sergei Borukhov,Ekaterina Stepanova,Jookyung Lee,Maria Ozerova,Ekaterina Semenova,Kirill Datsenko,Barry L Wanner,Konstantin Severinov,Sergei Borukhov,Ekaterina Stepanova,Jookyung Lee,Maria Ozerova,Ekaterina Semenova,Kirill Datsenko,Barry L Wanner,Konstantin Severinov,Sergei Borukhov,

Transcription elongation factor GreA induces nucleolytic activity of bacterial RNA polymerase (RNAP). In vitro, transcript cleavage by GreA contributes to transcription efficiency by (i) suppressing pauses and arrests, (ii) stimulating RNAP promoter escape, and (iii) enhancing transcription fidelity. However, it is unclear which of these functions is (are) most relevant in vivo. By comparing global gene expression profiles of Escherichia coli strains lacking Gre factors and strains expressing either the wild type (wt) or a functionally inactive GreA mutant, we identified genes that are potential targets of GreA action. Data analysis revealed that in the presence of chromosomally expressed GreA, 19 genes are upregulated; an additional 105 genes are activated upon overexpression of the wt but not the mutant GreA. Primer extension reactions with selected transcription units confirmed the gene array data. The most prominent stimulatory effect (threefold to about sixfold) of GreA was observed for genes of ribosomal protein operons and the tna operon, suggesting that transcript cleavage by GreA contributes to optimal expression levels of these genes in vivo. In vitro transcription assays indicated that the stimulatory effect of GreA upon the transcription of these genes is mostly due to increased RNAP recycling due to facilitated promoter escape. We propose that transcript cleavage during early stages of initiation is thus the main in vivo function of GreA. Surprisingly, the presence of the wt GreA also led to the decreased transcription of many genes. The mechanism of this effect is unknown and may be indirect.

Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro. Publishing Authors By Initials

E Stepanova,J Lee,M Ozerova,E Semenova,K Datsenko,BL Wanner,K Severinov,S Borukhov,E Stepanova,J Lee,M Ozerova,E Semenova,K Datsenko,BL Wanner,K Severinov,S Borukhov,E Stepanova,J Lee,M Ozerova,E Semenova,K Datsenko,BL Wanner,K Severinov,S Borukhov,

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Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Journal of bacteriology

VOLUME: 189

Page Numbers: 8772-85

Journal Abbreviation: J. Bacteriol.

ISSN: 1098-5530

DAY: 31

MONTH: 08

YEAR: 2007

Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro. Information

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LANGUAGE: eng

NlmUniqueID: 2985120

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Grant and Affiliation Information for Analysis of promoter targets for Escherichia coli transcription elongation factor GreA in vivo and in vitro.

AFFILIATION: Department of Cell Biology, UMDNJ-SOM at Stratford, 2 Medical Center Drive, Stratford, NJ 08084-1489, USA.

Country: United States

AGENCY: United States NIGMS

GRANT: GM62662

ACRONYM: GM

MEDLINETA: J Bacteriol

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