HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 43, 36397-36408, October 28, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/43/36397    most recent M507146200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Greive, S. J. Articles by von Hippel, P. H. Search for Related Content PubMed PubMed Citation Articles by Greive, S. J. Articles by von Hippel, P. H. Social Bookmarking                      What's this?

Assembly of an RNA-Protein Complex

BINDING OF NusB AND NusE (S10) PROTEINS TO boxA RNA NUCLEATES THE FORMATION OF THE ANTITERMINATION COMPLEX INVOLVED IN CONTROLLING rRNA TRANSCRIPTION IN ESCHERICHIA COLI^*

From the Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403

Analytical ultracentrifugation and fluorescence anisotropy methods have been used to measure the equilibrium parameters that control the formation of the core subcomplex of NusB and NusE proteins and boxA RNA. This subcomplex, in turn, nucleates the assembly of the antitermination complex that is involved in controlling the synthesis of ribosomal RNA in Escherichia coli and that also participates in forming the N protein-dependent antitermination complex in lambdoid phage synthesis. In this study we determined the dissociation constants (K_d values) for the individual binary interactions that participate in the assembly of the ternary NusB-NusE-boxA RNA subassembly, and we showed that multiple equilibria, involving both specific and nonspecific binding, are involved in the assembly pathway of this protein-RNA complex. The measured K_d values were used to model the in vitro assembly reaction and combined with in vivo concentration data to simulate the overall control of the assembly of this complex in E. coli at two different cellular growth rates. The results showed that at both growth rates assembly proceeds via the initial formation of a weak but specific NusB-boxA complex, which is then stabilized by NusE binding. We showed that NusE also binds nonspecifically to available single-stranded RNA sequences and that such nonspecific protein binding to RNA can help to regulate crucial interactions in the assembly of the various macromolecular machines of gene expression.

Received for publication, June 30, 2005 , and in revised form, August 17, 2005.

^* This work was supported by National Institutes of Health Grants GM-15792 and GM-29158 (to P. H. v. H.), by American Heart Association Postdoctoral Fellowship 0425713Z (to S. J. G.), and by an American Cancer Society Research Professorship (to P. H. v. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Present address: Dept. of Cell and Molecular Biology, University of Hawaii, Manoa, 1960 East-West Rd., Biomed. T-514, Honolulu, HI 96822.

² To whom correspondence should be addressed: Institute of Molecular Biology and Dept. of Chemistry, University of Oregon, Eugene, OR 97403. E-mail: petevh{at}molbio.uoregon.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S. Prasch, M. Jurk, R. S. Washburn, M. E. Gottesman, B. M. Wohrl, and P. Rosch RNA-binding specificity of E. coli NusA Nucleic Acids Res., June 10, 2009; (2009) gkp452v1. [Abstract] [Full Text] [PDF]

				K. B. Arnvig, S. Zeng, S. Quan, A. Papageorge, N. Zhang, A. C. Villapakkam, and C. L. Squires Evolutionary Comparison of Ribosomal Operon Antitermination Function J. Bacteriol., November 1, 2008; 190(21): 7251 - 7257. [Abstract] [Full Text] [PDF]

				P. J. Lewis, G. P. Doherty, and J. Clarke Transcription factor dynamics Microbiology, July 1, 2008; 154(7): 1837 - 1844. [Abstract] [Full Text] [PDF]

				G. P. Doherty, D. H. Meredith, and P. J. Lewis Subcellular Partitioning of Transcription Factors in Bacillus subtilis J. Bacteriol., June 1, 2006; 188(11): 4101 - 4110. [Abstract] [Full Text] [PDF]