Solution structure of YaeO, a rho-specific inhibitor of transcription termination

Pablo Gutiérrez, Guennadi Kozlov, Lisa Gabrielli, Demetra Elias, Michael J. Osborne, Imed Gallouzi, Kalle Gehring

Research output: Contribution to journalArticle

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Abstract

Rho-dependent transcription termination is an essential process for the regulation of bacterial gene expression. Thus far, only two Rho-specific inhibitors of bacterial transcription termination have been described, the psu protein from the satellite bacteriophage P4 and YaeO from Escherichia coli. Here, we report the solution structure of YaeO, the first of a Rho-specific inhibitor of transcription termination. YaeO is an acidic protein composed of an N-terminal helix and a seven-stranded β sandwich. NMR chemical shift perturbation experiments revealed that YaeO binds proximal to the primary nucleic acid binding site of Rho. Based on the NMR titrations, a docked model of the YaeO-Rho complex was calculated. These results suggest that YaeO binds outside the Rho hexamer, acting as a competitive inhibitor of RNA binding. In vitro gel shift assays confirmed the inhibition of nucleic acid binding to Rho. Site-directed mutagenesis showed that the negative character of YaeO is essential for its function in vivo.

Original languageEnglish
Pages (from-to)23348-23353
Number of pages6
JournalJournal of Biological Chemistry
Volume282
Issue number32
DOIs
Publication statusPublished - 10 Aug 2007
Externally publishedYes

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Transcription
Bacterial Gene Expression Regulation
Nucleic Acids
Site-Directed Mutagenesis
Nuclear magnetic resonance
Bacteriophages
Mutagenesis
Proteins
Gels
Binding Sites
Chemical shift
RNA
Escherichia coli
Titration
Gene expression
Assays
Satellites
Experiments
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Gutiérrez, P., Kozlov, G., Gabrielli, L., Elias, D., Osborne, M. J., Gallouzi, I., & Gehring, K. (2007). Solution structure of YaeO, a rho-specific inhibitor of transcription termination. Journal of Biological Chemistry, 282(32), 23348-23353. https://doi.org/10.1074/jbc.M702010200

Solution structure of YaeO, a rho-specific inhibitor of transcription termination. / Gutiérrez, Pablo; Kozlov, Guennadi; Gabrielli, Lisa; Elias, Demetra; Osborne, Michael J.; Gallouzi, Imed; Gehring, Kalle.

In: Journal of Biological Chemistry, Vol. 282, No. 32, 10.08.2007, p. 23348-23353.

Research output: Contribution to journalArticle

Gutiérrez, P, Kozlov, G, Gabrielli, L, Elias, D, Osborne, MJ, Gallouzi, I & Gehring, K 2007, 'Solution structure of YaeO, a rho-specific inhibitor of transcription termination', Journal of Biological Chemistry, vol. 282, no. 32, pp. 23348-23353. https://doi.org/10.1074/jbc.M702010200
Gutiérrez, Pablo ; Kozlov, Guennadi ; Gabrielli, Lisa ; Elias, Demetra ; Osborne, Michael J. ; Gallouzi, Imed ; Gehring, Kalle. / Solution structure of YaeO, a rho-specific inhibitor of transcription termination. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 32. pp. 23348-23353.
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