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Journal of Bacteriology, May 2006, p. 3674-3681, Vol. 188, No. 10
0021-9193/06/$08.00+0     doi:10.1128/JB.188.10.3674-3681.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of a Mutation in the Bacillus subtilis S-Adenosylmethionine Synthetase Gene That Results in Derepression of S-Box Gene Expression

Brooke A. McDaniel, Frank J. Grundy, Vineeta P. Kurlekar, Jerneja Tomsic, and Tina M. Henkin*

Department of Microbiology, The Ohio State University, Columbus, Ohio 43210

Received 7 October 2005/ Accepted 25 February 2006

Genes in the S-box family are regulated by binding of S-adenosylmethionine (SAM) to the 5' region of the mRNA of the regulated gene. SAM binding was previously shown to promote a rearrangement of the RNA structure that results in premature termination of transcription in vitro and repression of expression of the downstream coding sequence. The S-box RNA element therefore acts as a SAM-binding riboswitch in vitro. In an effort to identify factors other than SAM that could be involved in the S-box regulatory mechanism in vivo, we searched for trans-acting mutations in Bacillus subtilis that act to disrupt repression of S-box gene expression during growth under conditions where SAM pools are elevated. We identified a single mutant that proved to have one nucleotide substitution in the metK gene, encoding SAM synthetase. This mutation, designated metK10, resulted in a 15-fold decrease in SAM synthetase activity and a 4-fold decrease in SAM concentration in vivo. The metK10 mutation specifically affected S-box gene expression, and the increase in expression under repressing conditions was dependent on the presence of a functional transcriptional antiterminator element. The observation that the mutation identified in this search affects SAM production supports the model that the S-box RNAs directly monitor SAM in vivo, without a requirement for additional factors.

* Corresponding author. Mailing address: Department of Microbiology, The Ohio State University, 484 W. 12th Ave., Columbus, OH 43210. Phone: (614) 688-3831. Fax: (614) 292-8120. E-mail: henkin.3{at}osu.edu.

Journal of Bacteriology, May 2006, p. 3674-3681, Vol. 188, No. 10
0021-9193/06/$08.00+0     doi:10.1128/JB.188.10.3674-3681.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tomsic, J., McDaniel, B. A., Grundy, F. J., Henkin, T. M. (2008). Natural Variability in S-Adenosylmethionine (SAM)-Dependent Riboswitches: S-Box Elements in Bacillus subtilis Exhibit Differential Sensitivity to SAM In Vivo and In Vitro. J. Bacteriol. 190: 823-833 [Abstract] [Full Text]  
  • Sperandio, B., Gautier, C., McGovern, S., Ehrlich, D. S., Renault, P., Martin-Verstraete, I., Guedon, E. (2007). Control of Methionine Synthesis and Uptake by MetR and Homocysteine in Streptococcus mutans. J. Bacteriol. 189: 7032-7044 [Abstract] [Full Text]  
  • Fuchs, R. T., Grundy, F. J., Henkin, T. M. (2007). S-adenosylmethionine directly inhibits binding of 30S ribosomal subunits to the SMK box translational riboswitch RNA. Proc. Natl. Acad. Sci. USA 104: 4876-4880 [Abstract] [Full Text]  
  • HENKIN, T.M., GRUNDY, F.J. (2006). Sensing Metabolic Signals with Nascent RNA Transcripts: The T Box and S Box Riboswitches as Paradigms. Cold Spring Harb Symp Quant Biol 71: 231-237 [Abstract]  


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