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Journal of Bacteriology, January 2003, p. 107-114, Vol. 185, No. 1
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.1.107-114.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Inactivation of the selB Gene in Methanococcus maripaludis: Effect on Synthesis of Selenoproteins and Their Sulfur-Containing Homologs

Michael Rother,1,{dagger} Isabella Mathes,2 Friedrich Lottspeich,2 and August Böck1*

Microbiology, Department of Biology I, University of Munich, D-80638 Munich,1 Department of Protein Chemistry, Max-Planck-Institute of Biochemistry, D-82152 Martinsried, Germany2

Received 9 August 2002/ Accepted 1 October 2002

The genome of Methanococcus maripaludis harbors genes for at least six selenocysteine-containing proteins and also for homologs that contain a cysteine codon in the position of the UGA selenocysteine codon. To investigate the synthesis and function of both the Se and the S forms, a mutant with an inactivated selB gene was constructed and analyzed. The mutant was unable to synthesize any of the selenoproteins, thus proving that the gene product is the archaeal translation factor (aSelB) specialized for selenocysteine insertion. The wild-type form of M. maripaludis repressed the synthesis of the S forms of selenoproteins, i.e., the selenium-independent alternative system, in selenium-enriched medium, but the mutant did not. We concluded that free selenium is not involved in regulation but rather a successional compound such as selenocysteyl-tRNA or some selenoprotein. Apart from the S forms, several enzymes from the general methanogenic route were affected by selenium supplementation of the wild type or by the selB mutation. Although the growth of M. maripaludis on H2/CO2 is only marginally affected by the selB lesion, the gene is indispensable for growth on formate because M. maripaludis possesses only a selenocysteine-containing formate dehydrogenase.

* Corresponding author. Mailing address: Mikrobiologie, Department für Biologie I, Maria-Ward-Strasse 1a, D-80638 Munich, Germany. Phone: 49(0)89-2180-6120. Fax: 49(0)89-2180-6122. E-mail: august.boeck{at}lrz.uni-muenchen.de.

{dagger} Present address: Department of Microbiology, University of Illinois, Urbana, IL 61801.

Journal of Bacteriology, January 2003, p. 107-114, Vol. 185, No. 1
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.1.107-114.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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