This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Enoru-Eta, J.
Right arrow Articles by Charlier, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Enoru-Eta, J.
Right arrow Articles by Charlier, D.

 Previous Article  |  Next Article 

Journal of Bacteriology, July 2000, p. 3661-3672, Vol. 182, No. 13
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Purification and Characterization of Sa-Lrp, a DNA-Binding Protein from the Extreme Thermoacidophilic Archaeon Sulfolobus acidocaldarius Homologous to the Bacterial Global Transcriptional Regulator Lrp

Julius Enoru-Eta,1 Daniel Gigot,2 Thia-Lin Thia-Toong,1 Nicolas Glansdorff,1,2 and Daniel Charlier1,*

Erfelijkheidsleer en Microbiologie, Vrije Universiteit Brussel, and Department of Microbiology, The Flanders Interuniversity Institute for Biotechnology,1 and Laboratoire de Microbiologie, Université Libre de Bruxelles, and Institut de Recherches Microbiologiques J.-M. Wiame,2 B-1070 Brussels, Belgium

Received 3 February 2000/Accepted 10 April 2000

Archaea, constituting the third primary domain of life, harbor a basal transcription apparatus of the eukaryotic type, whereas curiously, a large fraction of the potential transcription regulation factors appear to be of the bacterial type. To date, little information is available on these predicted regulators and on the intriguing interplay that necessarily has to occur with the transcription machinery. Here, we focus on Sa-lrp of the extremely thermoacidophilic crenarchaeote Sulfolobus acidocaldarius, encoding an archaeal homologue of the Escherichia coli leucine-responsive regulatory protein Lrp, a global transcriptional regulator and genome organizer. Sa-lrp was shown to produce a monocistronic mRNA that was more abundant in the stationary-growth phase and produced in smaller amounts in complex medium, this down regulation being leucine independent. We report on Sa-Lrp protein purification from S. acidocaldarius and from recombinant E. coli, both identified by N-terminal amino acid sequence determination. Recombinant Sa-Lrp was shown to be homotetrameric and to bind to its own control region; this binding proved to be leucine independent and was stimulated at high temperatures. Interference binding experiments suggested an important role for minor groove recognition in the Sa-Lrp-DNA complex formation, and mutant analysis indicated the importance for DNA binding of the potential helix-turn-helix motif present at the N terminus of Sa-Lrp. The DNA-binding capacity of purified Sa-Lrp was found to be more resistant to irreversible heat inactivation in the presence of L-leucine, suggesting a potential physiological role of the amino acid as a cofactor.

* Corresponding author. Mailing address: Erfelijkheidsleer en Microbiologie, Vrije Universiteit Brussel, 1-av. E. Gryson, B-1070 Brussels, Belgium. Phone: 32 2 526 72 79. Fax: 32 2 526 72 73. E-mail: dcharlie{at}vub.ac.be.

Journal of Bacteriology, July 2000, p. 3661-3672, Vol. 182, No. 13
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Peeters, E., Wartel, C., Maes, D., Charlier, D. (2007). Analysis of the DNA-binding sequence specificity of the archaeal transcriptional regulator Ss-LrpB from Sulfolobus solfataricus by systematic mutagenesis and high resolution contact probing. Nucleic Acids Res 35: 623-633 [Abstract] [Full Text]  
  • Chen, L., Brugger, K., Skovgaard, M., Redder, P., She, Q., Torarinsson, E., Greve, B., Awayez, M., Zibat, A., Klenk, H.-P., Garrett, R. A. (2005). The Genome of Sulfolobus acidocaldarius, a Model Organism of the Crenarchaeota. J. Bacteriol. 187: 4992-4999 [Abstract] [Full Text]  
  • Gregor, D., Pfeifer, F. (2005). In vivo analyses of constitutive and regulated promoters in halophilic archaea. Microbiology 151: 25-33 [Abstract] [Full Text]  
  • Hofacker, A., Schmitz, K.-M., Cichonczyk, A., Sartorius-Neef, S., Pfeifer, F. (2004). GvpE- and GvpD-mediated transcription regulation of the p-gvp genes encoding gas vesicles in Halobacterium salinarum. Microbiology 150: 1829-1838 [Abstract] [Full Text]  
  • Lee, S.-J., Engelmann, A., Horlacher, R., Qu, Q., Vierke, G., Hebbeln, C., Thomm, M., Boos, W. (2003). TrmB, a Sugar-specific Transcriptional Regulator of the Trehalose/Maltose ABC Transporter from the Hyperthermophilic Archaeon Thermococcus litoralis. J. Biol. Chem. 278: 983-990 [Abstract] [Full Text]  
  • Thia-Toong, T.-L., Roovers, M., Durbecq, V., Gigot, D., Glansdorff, N., Charlier, D. (2002). Genes of De Novo Pyrimidine Biosynthesis from the Hyperthermoacidophilic Crenarchaeote Sulfolobus acidocaldarius: Novel Organization in a Bipolar Operon. J. Bacteriol. 184: 4430-4441 [Abstract] [Full Text]  
  • Brinkman, A. B., Bell, S. D., Lebbink, R. J., de Vos, W. M., van der Oost, J. (2002). The Sulfolobus solfataricus Lrp-like Protein LysM Regulates Lysine Biosynthesis in Response to Lysine Availability. J. Biol. Chem. 277: 29537-29549 [Abstract] [Full Text]  
  • Dahlke, I., Thomm, M. (2002). A Pyrococcus homolog of the leucine-responsive regulatory protein, LrpA, inhibits transcription by abrogating RNA polymerase recruitment. Nucleic Acids Res 30: 701-710 [Abstract] [Full Text]  
  • Napoli, A., Kvaratskelia, M., White, M. F., Rossi, M., Ciaramella, M. (2001). A Novel Member of the Bacterial-Archaeal Regulator Family Is a Nonspecific DNA-binding Protein and Induces Positive Supercoiling. J. Biol. Chem. 276: 10745-10752 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»