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Microbiology 147 (2001), 153-159
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Microbiology (2001), 147, 153-159.
© 2001 Society for General Microbiology

Genetics and Molecular Biology

Anomalous DnaA protein binding to the regulatory region of the Escherichia coli aldA gene

Toru Ozaki1, Yuichi Kumakia,1, Risa Kitagawab,1 and Tohru Ogawa1

Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan1

Author for correspondence: Tohru Ogawa. Tel: +81 52 789 2584. Fax: +81 52 789 5732. e-mail: h44851a{at}nucc.cc.nagoya-u.ac.jp

A binding site for DnaA protein was identified in the regulatory region of the aldA gene of Escherichia coli. Specific binding was demonstrated by in vitro assays including filter binding as well as mobility shift in a polyacrylamide gel of the DnaA–DNA complex. In cells growing in minimal medium containing glucose, expression of ß-galactosidase activity from an aldA–lacZ fusion gene was suppressed by oversupply of DnaA protein and was enhanced by reducing the free DnaA level. These results suggest that DnaA protein negatively regulates expression of the aldA gene under these conditions. Despite fairly strong binding, the bound DNA fragment had no consensus 9 bp DnaA binding sequence (DnaA box), and anomalous binding to an AT-rich sequence located close to the transcription start site was revealed by a footprinting experiment.

Keywords: DnaA, aldA, transcription, replication, Escherichia coli

a Present address: INTEC Web and Genome Informatics Corporation, Bio Informatics Group, 1-3-3 Shinsuna, Koto-ku, Tokyo 136-0075, Japan.

b Present address: Department of Medical Genetics, University of British Columbia, 6174 University Boulevard, Vancouver, BC, Canada V6T 1Z3.






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Copyright © 2001 Society for General Microbiology.