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Journal of Bacteriology, January 2000, p. 252-255, Vol. 182, No. 1
0021-9193/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A celR Mutation Affecting Transcription of Cellulase Genes in Thermobifida fusca

Nikolay A. Spiridonov and David B. Wilson*

Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853

Received 25 August 1999/Accepted 8 October 1999

Biosynthesis of extracellular cellulases in the cellulose-degrading actinomycete Thermobifida fusca is controlled by a transcriptional regulator, CelR, and cellobiose, which acts as an inducer interfering with the CelR-DNA interaction. We report the identification and characterization of a mutation in the celR gene that changes Ala55 in the hinge helix of CelR to Thr. The wild-type and mutant celR genes were cloned in Escherichia coli, and their protein products were characterized. The CelR mutant protein bound DNA more weakly than the wild-type protein and formed a less stable complex with DNA in the presence of cellobiose. The results of Western analysis and gel retardation experiments suggest that CelR is produced constitutively and its DNA-binding activity is regulated through posttranslational modification.

* Corresponding author. Mailing address: 458 Biotechnology Bldg., Cornell University, Ithaca, NY 14853. Phone: (607) 255-5706. Fax: (607) 255-2428. E-mail: dbw3{at}cornell.edu.

Journal of Bacteriology, January 2000, p. 252-255, Vol. 182, No. 1
0021-9193/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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