Multiple interactions with DNA, RNA and transcription factors occur in a transcription cycle. To survey the proximity of some of these factors to the Escherichiacoli RNA polymerase surface, we produced a set of nine monoclonal antibodies (mAbs) against the enzyme. These mAbs, located at different places on the surface of the enzyme, were used in a co-immunopurification assay to investigate interference with the binding of NusA, σ70, GreB and HepA to core RNA polymerase. One of these mAbs turned out to be the first antibody inhibitor of the binding of NusA and σ70; it did not affect GreB and HepA interactions. Its epitope was located on the β′ subunit at the C-terminus of region G. The properties of this mAb reinforce the idea that the mutually exclusive binding of NusA and σ70 to core RNA polymerase is due to, at least partially, overlapping binding sites, rather than allosteric interaction between two distant binding sites. This mAb is also useful to understand the occupancy of σ70, NusA and Gre proteins on core RNA polymerase.
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January 2002
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Research Article|
January 08 2002
Characterization of monoclonal antibodies against Escherichia coli core RNA polymerase
Jöelle ROUBY;
Jöelle ROUBY
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
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Martine PUGNIERE;
Martine PUGNIERE
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
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Jean-Claude MANI;
Jean-Claude MANI
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
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Claude GRANIER;
Claude GRANIER
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
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Pierrette MONMOUTON;
Pierrette MONMOUTON
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
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Stephane THEULIER SAINT GERMAIN;
Stephane THEULIER SAINT GERMAIN
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
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Jean-Paul LEONETTI
Jean-Paul LEONETTI
1
Centre National de la Recherche Scietifique (CNRS), Unité Mixte de Recherche (UMR) 5094, Institut de Biotechnologie et Pharmacologie, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France
1To whom correspondence should be addressed (e-mail jp_leo@yahoo.com).
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Publisher: Portland Press Ltd
Received:
May 14 2001
Revision Received:
October 09 2001
Accepted:
November 05 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 361 (2): 347–354.
Article history
Received:
May 14 2001
Revision Received:
October 09 2001
Accepted:
November 05 2001
Citation
Jöelle ROUBY, Martine PUGNIERE, Jean-Claude MANI, Claude GRANIER, Pierrette MONMOUTON, Stephane THEULIER SAINT GERMAIN, Jean-Paul LEONETTI; Characterization of monoclonal antibodies against Escherichia coli core RNA polymerase. Biochem J 15 January 2002; 361 (2): 347–354. doi: https://doi.org/10.1042/bj3610347
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