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Originally published In Press as doi:10.1074/jbc.M000390200 on June 8, 2000

J. Biol. Chem., Vol. 275, Issue 37, 29061-29065, September 15, 2000
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Role of Activating Region 1 of Escherichia coli FNR Protein in Transcription Activation at Class II Promoters*

Helen J. WingDagger §, Jeff Green, John R. Guest, and Stephen J. W. BusbyDagger

From the Dagger  School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom and the  Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, United Kingdom

FNR is an Escherichia coli transcription factor that activates gene expression in response to anaerobiosis at a large number of promoters by making direct contacts with RNA polymerase. At class II FNR-dependent promoters, where the DNA site for FNR overlaps the -35 element, activating region 1 of FNR is proposed to interact with the C-terminal domain of the RNA polymerase alpha -subunit. Using a model class II FNR-dependent promoter, FF(-41.5), we have performed in vivo and in vitro experiments to investigate the role of this interaction. Our results show that FNR, carrying substitutions in activating region 1, is compromised in its ability to promote open complex formation and thus to activate transcription. Abortive initiation assays were used to assess the contribution of activating region 1 of FNR to open complex formation. A new method for the purification of the FNR protein is also described.

* This work was funded by a Biotechnology and Biological Science Research Council project Grant G06544.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed. Present address: Harvard School of Public Health, 665 Huntington Ave., Dept. I.I.D., Bldg. 1, Rm. 902, Boston, MA 02115. Tel.: 617-432-3239; Fax: 617-432-3259; E-mail: hwing@hsph.harvard.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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