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Journal of Bacteriology, May 2000, p. 2771-2777, Vol. 182, No. 10
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The Bacillus subtilis GTP Binding Protein Obg and Regulators of the sigma B Stress Response Transcription Factor Cofractionate with Ribosomes

Janelle M. Scott, Jingliang Ju,dagger Theresa Mitchell, and W. G. Haldenwang*

Department of Microbiology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900

Received 30 November 1999/Accepted 15 February 2000

Obg, an essential GTP binding protein of Bacillus subtilis, is necessary for stress activation of the sigma B transcription factor. We investigated Obg's cellular associations by differential centrifugation of crude B. subtilis extracts, using an anti-Obg antibody as a probe to monitor Obg during the fractionation, and by fluorescent microscopy of a B. subtilis strain in which Obg was fused to green fluorescent protein. The results indicated that Obg is part of a large cytoplasmic complex. In subsequent analyses, Obg coeluted with ribosomal subunits during gel filtration of B. subtilis lysates on Sephacryl S-400 and specifically bound to ribosomal protein L13 in an affinity blot assay. Probing the gel filtration fractions with antibodies specific for sigma B and its coexpressed regulators (Rsb proteins) revealed coincident elution of the upstream components of the sigma B stress activation pathway (RsbR, -S, and -T) with Obg and the ribosomal subunits. The data implicate ribosome function as a possible mediator of the activity of Obg and the stress induction of sigma B.

* Corresponding author. Mailing address: Dept. of Microbiology, UTHSCSA, 7703 Floyd Curl Dr., MC 7758, San Antonio, TX 78229-3900. Phone: (210) 567-3957. Fax: (210) 567-6612. E-mail: haldenwang{at}uthscsa.edu.

dagger Present address: Department of Food Science and Technology, University of Nebraska, Lincoln, NE 68585-0919.

Journal of Bacteriology, May 2000, p. 2771-2777, Vol. 182, No. 10
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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