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

J. Biol. Chem., Vol. 281, Issue 46, 34909-34917, November 17, 2006

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 281/46/34909    most recent M607582200v1 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 Heinrich, A. Articles by Forchhammer, K. Search for Related Content PubMed PubMed Citation Articles by Heinrich, A. Articles by Forchhammer, K. Social Bookmarking                      What's this?

Interaction of the Membrane-bound GlnK-AmtB Complex with the Master Regulator of Nitrogen Metabolism TnrA in Bacillus subtilis^*

Annette Heinrich, Kathrin Woyda, Katja Brauburger, Gregor Meiss, Christian Detsch^¶, Jörg Stülke^||, and Karl Forchhammer¹

From the Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany, Institut für Biochemie Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 58, D-35392 Giessen, Germany, ^¶Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, D-91058 Erlangen, Germany, and ^||Institut für Mikrobiologie und Genetik, Georg-August Universität Göttingen, Grisebachstrasse 8, D-37077 Göttingen, Germany

P_II proteins are widespread and highly conserved signal transduction proteins occurring in bacteria, Archaea, and plants and play pivotal roles in controlling nitrogen assimilatory metabolism. This study reports on biochemical properties of the P_II-homologue GlnK (originally termed NrgB) in Bacillus subtilis (BsGlnK). Like other P_II proteins, the native BsGlnK protein has a trimeric structure and readily binds ATP in the absence of divalent cations, whereas 2-oxoglutarate is only weakly bound. In contrast to other P_II-like proteins, Mg²⁺ severely affects its ATP-binding properties. BsGlnK forms a tight complex with the membrane-bound ammonium transporter AmtB (NrgA), from which it can be relieved by millimolar concentrations of ATP. Immunoprecipitation and co-localization experiments identified a novel interaction between the BsGlnK-AmtB complex and the major transcription factor of nitrogen metabolism, TnrA. In vitro in the absence of ATP, TnrA is completely tethered to membrane (AmtB)-bound GlnK, whereas in extracts from BsGlnK- or AmtB-deficient cells, TnrA is entirely soluble. The presence of 4 mM ATP leads to concomitant solubilization of BsGlnK and TnrA. This ATP-dependent membrane re-localization of TnrA by BsGlnK/AmtB may present a novel mechanism to control the global nitrogen-responsive transcription regulator TnrA in B. subtilis under certain physiological conditions.

Received for publication, August 9, 2006 , and in revised form, September 20, 2006.

^* This work was supported by Deutsche Forschungsgemeinschaft Grant Fo 195/4. 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.

The on-line version of this article (available at http://www.jbc.org) contains Construction of Plasmids and detailed Purification Protocols and Refs. 1–6 and 60 and 61.

¹ To whom correspondence should be addressed. Tel.: 49-641-9935545; Fax: 49-641-9935549; E-mail: Karl.Forchhammer{at}mikro.bio.uni-giessen.de.

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

This article has been cited by other articles:

				P. F. Teixeira, A. Jonsson, M. Frank, H. Wang, and S. Nordlund Interaction of the signal transduction protein GlnJ with the cellular targets AmtB1, GlnE and GlnD in Rhodospirillum rubrum: dependence on manganese, 2-oxoglutarate and the ADP/ATP ratio Microbiology, August 1, 2008; 154(8): 2336 - 2347. [Abstract] [Full Text] [PDF]

				A. Kayumov, A. Heinrich, M. Sharipova, O. Iljinskaya, and K. Forchhammer Inactivation of the general transcription factor TnrA in Bacillus subtilis by proteolysis Microbiology, August 1, 2008; 154(8): 2348 - 2355. [Abstract] [Full Text] [PDF]

				D. M. Wolfe, Y. Zhang, and G. P. Roberts Specificity and Regulation of Interaction between the PII and AmtB1 Proteins in Rhodospirillum rubrum J. Bacteriol., October 1, 2007; 189(19): 6861 - 6869. [Abstract] [Full Text] [PDF]

				P.-L. Tremblay, T. Drepper, B. Masepohl, and P. C. Hallenbeck Membrane Sequestration of PII Proteins and Nitrogenase Regulation in the Photosynthetic Bacterium Rhodobacter capsulatus J. Bacteriol., August 15, 2007; 189(16): 5850 - 5859. [Abstract] [Full Text] [PDF]

				M. J. Conroy, A. Durand, D. Lupo, X.-D. Li, P. A. Bullough, F. K. Winkler, and M. Merrick The crystal structure of the Escherichia coli AmtB-GlnK complex reveals how GlnK regulates the ammonia channel PNAS, January 23, 2007; 104(4): 1213 - 1218. [Abstract] [Full Text] [PDF]