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Journal of Bacteriology, February 2001, p. 882-889, Vol. 183, No. 3
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.3.882-889.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Ammonia Switch-Off of Nitrogen Fixation in the Methanogenic Archaeon Methanococcus maripaludis: Mechanistic Features and Requirement for the Novel GlnB Homologues, NifI1 and NifI2

Peter S. Kessler,dagger Catherine Daniel, and John A. Leigh*

Department of Microbiology, University of Washington, Seattle, Washington 98195

Received 10 July 2000/Accepted 7 November 2000

Ammonia switch-off is the immediate inactivation of nitrogen fixation that occurs when a superior nitrogen source is encountered. In certain bacteria switch-off occurs by reversible covalent ADP-ribosylation of the dinitrogenase reductase protein, NifH. Ammonia switch-off occurs in diazotrophic species of the methanogenic Archaea as well. We showed previously that in Methanococcus maripaludis switch-off requires at least one of two novel homologues of glnB, a family of genes whose products play a central role in nitrogen sensing and regulation in bacteria. The novel glnB homologues have recently been named nifI1 and nifI2. Here we use in-frame deletions and genetic complementation analysis in M. maripaludis to show that the nifI1 and nifI2 genes are both required for switch-off. We could not detect ADP-ribosylation or any other covalent modification of dinitrogenase reductase during switch-off, suggesting that the mechanism differs from the well-studied bacterial system. Furthermore, switch-off did not affect nif gene transcription, nifH mRNA stability, or NifH protein stability. Nitrogenase activity resumed within a short time after ammonia was removed from a switched-off culture, suggesting that whatever the mechanism, it is reversible. We demonstrate the physiological importance of switch-off by showing that it allows growth to accelerate substantially when a diazotrophic culture is switched to ammonia.

* Corresponding author. Mailing address: University of Washington, Department of Microbiology, Box 357242, Seattle, WA 98195-7242. Phone: (206) 685-1390. Fax: (206) 543-8297. E-mail: leighj{at}u.washington.edu.

dagger Present address: Seattle Biomedical Research Institute, Seattle, WA 98195.

Journal of Bacteriology, February 2001, p. 882-889, Vol. 183, No. 3
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.3.882-889.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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