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Journal of Bacteriology, July 2006, p. 5167-5176, Vol. 188, No. 14
0021-9193/06/$08.00+0     doi:10.1128/JB.00318-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Regulation of the Myxococcus xanthus C-Signal-Dependent {Omega}4400 Promoter by the Essential Developmental Protein FruA

Deborah R. Yoder-Himes and Lee Kroos*

Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824

Received 3 March 2006/ Accepted 5 May 2006

The bacterium Myxococcus xanthus employs extracellular signals to coordinate aggregation and sporulation during multicellular development. Extracellular, contact-dependent signaling that involves the CsgA protein (called C-signaling) activates FruA, a putative response regulator that governs a branched signaling pathway inside cells. One branch regulates cell movement, leading to aggregation. The other branch regulates gene expression, leading to sporulation. C-signaling is required for full expression of most genes induced after 6 h into development, including the gene identified by Tn5 lac insertion {Omega}4400. To determine if FruA is a direct regulator of {Omega}4400 transcription, a combination of in vivo and in vitro experiments was performed. {Omega}4400 expression was abolished in a fruA mutant. The DNA-binding domain of FruA bound specifically to DNA upstream of the promoter –35 region in vitro. Mutations between bp –86 and –77 greatly reduced binding. One of these mutations had been shown previously to reduce {Omega}4400 expression in vivo and make it independent of C-signaling. For the first time, chromatin immunoprecipitation (ChIP) experiments were performed on M. xanthus. The ChIP experiments demonstrated that FruA is associated with the {Omega}4400 promoter region late in development, even in the absence of C-signaling. Based on these results, we propose that FruA directly activates {Omega}4400 transcription to a moderate level prior to C-signaling and, in response to C-signaling, binds near bp –80 and activates transcription to a higher level. Also, the highly localized effects of mutations between bp –86 and –77 on DNA binding in vitro, together with recently published footprints, allow us to predict a consensus binding site of GTCG/CGA/G for the FruA DNA-binding domain.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824. Phone: (517) 355-9726. Fax: (517) 353-9334. E-mail: kroos{at}msu.edu.

Journal of Bacteriology, July 2006, p. 5167-5176, Vol. 188, No. 14
0021-9193/06/$08.00+0     doi:10.1128/JB.00318-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Mittal, S., Kroos, L. (2009). Combinatorial Regulation by a Novel Arrangement of FruA and MrpC2 Transcription Factors during Myxococcus xanthus Development. J. Bacteriol. 191: 2753-2763 [Abstract] [Full Text]  
  • Mittal, S., Kroos, L. (2009). A combination of unusual transcription factors binds cooperatively to control Myxococcus xanthus developmental gene expression. Proc. Natl. Acad. Sci. USA 106: 1965-1970 [Abstract] [Full Text]  
  • Viswanathan, P., Murphy, K., Julien, B., Garza, A. G., Kroos, L. (2007). Regulation of dev, an Operon That Includes Genes Essential for Myxococcus xanthus Development and CRISPR-Associated Genes and Repeats. J. Bacteriol. 189: 3738-3750 [Abstract] [Full Text]  
  • Viswanathan, P., Ueki, T., Inouye, S., Kroos, L. (2007). Combinatorial regulation of genes essential for Myxococcus xanthus development involves a response regulator and a LysR-type regulator. Proc. Natl. Acad. Sci. USA 104: 7969-7974 [Abstract] [Full Text]  
  • Gronewold, T. M. A., Kaiser, D. (2007). Mutations of the Act Promoter in Myxococcus xanthus. J. Bacteriol. 189: 1836-1844 [Abstract] [Full Text]  


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