This Article Full Text Full Text (PDF) Supporting Information Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a colleague Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My File Cabinet Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via ISI Web of Science (7) Citing Articles via Google Scholar Google Scholar Articles by Bonin, I. Articles by Wahl, M. C. Search for Related Content PubMed PubMed Citation Articles by Bonin, I. Articles by Wahl, M. C. Pubmed/NCBI databases Gene GEO Profiles Protein Structure Social Bookmarking                What's this?

 Previous Article  | Table of Contents |  Next Article 

BIOCHEMISTRY
Structural basis for the interaction of Escherichia coli NusA with protein N of phage

Irena Bonin ^*, Rene Mühlberger , Gleb P. Bourenkov , Robert Huber ^*, Adelbert Bacher , Gerald Richter , and Markus C. Wahl ^, ¶

^*Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Am Klopferspitz 18a, D-82152 Martinsried, Germany; Technische Universität München, Institut für Organische Chemie und Biochemie, Lichtenbergstrasse 4, D-85747 Garching, Germany; Max-Planck-Arbeitsgruppen für Strukturelle Molekularbiologie, Deutsches Elektronen Synchrotron, Arbeitsgruppe Proteindynamik, Notkestrasse 85, D-22603 Hamburg, Germany; and Max-Planck-Institut für Biophysikalische Chemie, Abteilung Zelluläre Biochemie/Röntgenkristallographie, Am Fassberg 11, D-37077 Göttingen, Germany

Contributed by Robert Huber, August 11, 2004

The C terminus of transcription factor NusA from Escherichia coli comprises two repeat units, which bind during antitermination to protein N from phage . To delineate the structural basis of the NusA–N interaction, we attempted to crystallize the NusA C-terminal repeats in complex with a N peptide (residues 34–47). The two NusA domains became proteolytically separated during crystallization, and crystals contained two copies of the first repeat unit in contact with a single N fragment. The NusA modules employ identical regions to contact the peptide but approach the ligand from opposite sides. In contrast to the -helical conformation of the N N terminus in complex with boxB RNA, residues 34–40 of N remain extended upon interaction with NusA. Mutational analyses indicated that only one of the observed NusA–N interaction modes is biologically significant, supporting an equimolar ratio of NusA and N in antitermination complexes. Solution studies indicated that additional interactions are fostered by the second NusA repeat unit, consistent with known compensatory mutations in NusA and N. Contrary to the RNA polymerase subunit, N binding does not stimulate RNA interaction of NusA. The results demonstrate that N serves as a scaffold to closely oppose NusA and the mRNA in antitermination complexes.

Data deposition: The structure coordinates have been deposited in the Protein Data Bank, www.pdb.org (PDB ID code 1U9L).

^¶ To whom correspondence should be addressed. E-mail: mwahl{at}gwdg.de.

© 2004 by The National Academy of Sciences of the USA

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

This article has been cited by other articles in HighWire Press-hosted journals:

				A. Cheeran, N. R. Kolli, and R. Sen The Site of Action of the Antiterminator Protein N from the Lambdoid Phage H-19B J. Biol. Chem., October 19, 2007; 282(42): 30997 - 31007. [Abstract] [Full Text] [PDF]

				A. Eisenmann, S. Schwarz, S. Prasch, K. Schweimer, and P. Rosch The E. coli NusA carboxy-terminal domains are structurally similar and show specific RNAP- and {lambda}N interaction Protein Sci., August 1, 2005; 14(8): 2018 - 2029. [Abstract] [Full Text] [PDF]

Current Issue | Archives | Online Submission | Info for Authors | Editorial Board | About Subscribe | Advertise | Contact | Site Map Copyright © 2004 by the National Academy of Sciences