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Applied and Environmental Microbiology, April 2008, p. 2103-2110, Vol. 74, No. 7
0099-2240/08/$08.00+0     doi:10.1128/AEM.02855-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Transcription of hupSL in Anabaena variabilis ATCC 29413 Is Regulated by NtcA and Not by Hydrogen{triangledown}

Philip D. Weyman, Brenda Pratte, and Teresa Thiel*

University of Missouri—St. Louis, Department of Biology, Research 223, St. Louis, Missouri 63121

Received 18 December 2007/ Accepted 7 February 2008

Nitrogen-fixing cyanobacteria such as Anabaena variabilis ATCC 29413 use an uptake hydrogenase, encoded by hupSL, to recycle hydrogen gas that is produced as an obligate by-product of nitrogen fixation. The regulation of hupSL in A. variabilis is likely to differ from that of the closely related Anabaena sp. strain PCC 7120 because A. variabilis lacks the excision element-mediated regulation that characterizes hupSL regulation in strain PCC 7120. An analysis of the hupSL transcript in a nitrogenase mutant of A. variabilis that does not produce any detectable hydrogen indicated that neither nitrogen fixation nor hydrogen gas was required for the induction of hupSL. Furthermore, exogenous addition of hydrogen gas did not stimulate hupSL transcription. Transcriptional reporter constructs indicated that the accumulation of hupSL transcript after nitrogen step-down was restricted primarily to the microaerobic heterocysts. Anoxic conditions were not sufficient to induce hupSL transcription. The induction of hupSL after nitrogen step-down was reduced in a mutant in the global nitrogen regulator NtcA, but was not reduced in a mutant unable to form heterocysts. A consensus NtcA-binding site was identified upstream of hupSL, and NtcA was found to bind to this region. Thus, while neither hydrogen gas nor anoxia controlled the expression of hupSL, its expression was controlled by NtcA. Heterocyst differentiation was not required for hupSL induction in response to nitrogen step-down, but heterocyst-localized cues may add an additional level of regulation to hupSL.

* Corresponding author. Mailing address: University of Missouri—St. Louis, Department of Biology, Research 223, St. Louis, MO 63121. Phone: (314) 516-6208. Fax: (314) 516-6233. E-mail: thiel{at}umsl.edu

{triangledown} Published ahead of print on 15 February 2008.

Applied and Environmental Microbiology, April 2008, p. 2103-2110, Vol. 74, No. 7
0099-2240/08/$08.00+0     doi:10.1128/AEM.02855-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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