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

 Previous Article  |  Next Article 

J. Bacteriol., Sep 1996, 5159-5163, Vol 178, No. 17
Copyright © 1996, American Society for Microbiology

Interaction of the trp RNA-Binding attenuation protein (TRAP) of Bacillus subtilis with RNA: effects of the number of GAG repeats, the nucleotides separating adjacent repeats, and RNA secondary structure

P Babitzke, J Yealy and D Campanelli
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park 16802, USA.

The 11-subunit trp RNA-binding attenuation protein of Bacillus subtilis, TRAP, regulates transcription and translation by binding to several (G/U)AG repeats present in the trp leader and trpG transcripts. Filter binding assays were used to study interactions between L- tryptophan-activated TRAP and synthetic RNAs. RNAs that contained GAG and/or UAG repeats were tested while the length and sequence of the nucleotides separating adjacent trinucleotide repeats were altered. TRAP-RNA complexes formed with transcripts containing GAG repeats were more stable than those with transcripts containing UAG repeats or alternating GAG and UAG repeats. The stability of TRAP-RNA complexes also increased substantially when the number of GAG repeats was increased from five to six and from six to seven. A gradual increase in complex stability was observed when the number of GAG repeats was increased from 7 to 11. The optimal spacer between adjacent trinucleotide repeats was found to be 2 nucleotides, with A and U residues preferred over G and C residues. TRAP binding was specific for single-stranded RNA; TRAP could not bind to RNA containing GAG repeats base paired in a stable RNA duplex. Overall, our findings suggest that each L-tryptophan-activated TRAP subunit can bind one (G/U)AG repeat and that multiple TRAP subunit-RNA binding site interactions are required for stable TRAP-RNA association.

This article has been cited by other articles:

• McGraw, A. P., Mokdad, A., Major, F., Bevilacqua, P. C., Babitzke, P. (2009). Molecular basis of TRAP-5'SL RNA interaction in the Bacillus subtilis trp operon transcription attenuation mechanism. RNA 15: 55-66 [Abstract] [Full Text]  
• McGraw, A. P., Bevilacqua, P. C., Babitzke, P. (2007). TRAP-5' stem loop interaction increases the efficiency of transcription termination in the Bacillus subtilis trpEDCFBA operon leader region. RNA 13: 2020-2033 [Abstract] [Full Text]  
• Nie, M., Htun, H. (2006). Different modes and potencies of translational repression by sequence-specific RNA-protein interaction at the 5'-UTR. Nucleic Acids Res 34: 5528-5540 [Abstract] [Full Text]  
• Payal, V., Gollnick, P. (2006). Substitutions of Thr30 provide mechanistic insight into tryptophan-mediated activation of TRAP binding to RNA. Nucleic Acids Res 34: 2933-2942 [Abstract] [Full Text]  
• Szigeti, R., Milescu, M., Gollnick, P. (2004). Regulation of the Tryptophan Biosynthetic Genes in Bacillus halodurans: Common Elements but Different Strategies than Those Used by Bacillus subtilis. J. Bacteriol. 186: 818-828 [Abstract] [Full Text]  
• Yakhnin, H., Zhang, H., Yakhnin, A. V., Babitzke, P. (2004). The trp RNA-Binding Attenuation Protein of Bacillus subtilis Regulates Translation of the Tryptophan Transport Gene trpP (yhaG) by Blocking Ribosome Binding. J. Bacteriol. 186: 278-286 [Abstract] [Full Text]  
• SCHAAK, J. E., BABITZKE, P., BEVILACQUA, P. C. (2003). Phylogenetic conservation of RNA secondary and tertiary structure in the trpEDCFBA operon leader transcript in Bacillus. RNA 9: 1502-1515 [Abstract] [Full Text]  
• Li, P. T. X., Gollnick, P. (2002). Using Hetero-11-mers Composed of Wild Type and Mutant Subunits to Study Tryptophan Binding to TRAP and Its Role in Activating RNA Binding. J. Biol. Chem. 277: 35567-35573 [Abstract] [Full Text]  
• Li, P. T. X., Scott, D. J., Gollnick, P. (2002). Creating Hetero-11-mers Composed of Wild-type and Mutant Subunits to Study RNA Binding to TRAP. J. Biol. Chem. 277: 11838-11844 [Abstract] [Full Text]  
• Babitzke, P., Gollnick, P. (2001). Posttranscription Initiation Control of Tryptophan Metabolism in Bacillus subtilis by the trp RNA-Binding Attenuation Protein (TRAP), anti-TRAP, and RNA Structure. J. Bacteriol. 183: 5795-5802 [Full Text]  
• Du, H., Yakhnin, A. V., Dharmaraj, S., Babitzke, P. (2000). trp RNA-Binding Attenuation Protein-5' Stem-Loop RNA Interaction Is Required for Proper Transcription Attenuation Control of the Bacillus subtilis trpEDCFBA Operon. J. Bacteriol. 182: 1819-1827 [Abstract] [Full Text]  
• Sudershana, S., Du, H., Mahalanabis, M., Babitzke, P. (1999). A 5' RNA Stem-Loop Participates in the Transcription Attenuation Mechanism That Controls Expression of the Bacillus subtilis trpEDCFBA Operon. J. Bacteriol. 181: 5742-5749 [Abstract] [Full Text]  
• Xirasagar, S., Elliott, M. B., Bartolini, W., Gollnick, P., Gottlieb, P. A. (1998). RNA Structure Inhibits the TRAP (trp RNA-binding Attenuation Protein)-RNA Interaction. J. Biol. Chem. 273: 27146-27153 [Abstract] [Full Text]  
• Du and Paul Babitzke, H. (1998). trp RNA-binding Attenuation Protein-mediated Long Distance RNA Refolding Regulates Translation of trpE in Bacillus subtilis. J. Biol. Chem. 273: 20494-20503 [Abstract] [Full Text]  
• Baumann, C., Xirasagar, S., Gollnick, P. (1997). The trp RNA-binding Attenuation Protein (TRAP) from Bacillus subtilis Binds to Unstacked trp Leader RNA. J. Biol. Chem. 272: 19863-19869 [Abstract] [Full Text]