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Journal of Bacteriology, January 2004, p. 580-587, Vol. 186, No. 2
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.2.580-587.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Expression and Regulation of a Silent Operon, hyf, Coding for Hydrogenase 4 Isoenzyme in Escherichia coli{dagger}

William T. Self,1 Adnan Hasona,2 and K. T. Shanmugam2*

Department of Molecular Biology and Microbiology, University of Central Florida, Orlando, Florida 32816 ,1 Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida 326112

Received 24 June 2003/ Accepted 20 October 2003

On the basis of hyf-lacZ fusion studies, the hyf operon of Escherichia coli, noted for encoding the fourth hydrogenase isoenzyme (HYD4), is not expressed at a significant level in a wild-type strain. However, mutant FhlA proteins (constitutive activators of the hyc-encoded hydrogenase 3 isoenzyme) activated hyf-lacZ. HyfR, an FhlA homolog encoded by the hyfR gene present at the end of the hyf operon, also activated transcription of hyf-lacZ but did so only when hyfR was expressed from a heterologous promoter. The HYD4 isoenzyme did not substitute for HYD3 in H2 production. Optimum expression of hyf-lacZ required the presence of cyclic AMP receptor protein-cyclic AMP complex and anaerobic conditions when HyfR was the activator.

* Corresponding author. Mailing address: Department of Microbiology and Cell Science, Box 110700, University of Florida, Gainesville, FL 32611. Phone: (352) 392-2490. Fax: (352) 392-5922. E-mail: shan{at}ufl.edu.

{dagger} Florida Agricultural Experiment Station Journal Series no. R-09737.

Journal of Bacteriology, January 2004, p. 580-587, Vol. 186, No. 2
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.2.580-587.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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