Advertisement
JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 QUICK SEARCH:   [advanced]


     


Originally published In Press as doi:10.1074/jbc.M301992200 on May 20, 2003

J. Biol. Chem., Vol. 278, Issue 31, 28711-28718, August 1, 2003
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
278/31/28711    most recent
M301992200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Little, R.
Right arrow Articles by Dixon, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Little, R.
Right arrow Articles by Dixon, R.
Social Bookmarking
 Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

The Amino-terminal GAF Domain of Azotobacter vinelandii NifA Binds 2-Oxoglutarate to Resist Inhibition by NifL under Nitrogen-limiting Conditions*

Richard Little and Ray Dixon {ddagger}

From the Department of Molecular Microbiology, John Innes Centre, Norwich NR4 7UH, United Kingdom

The expression of genes required for the synthesis of molybdenum nitrogenase in Azotobacter vinelandii is controlled by the NifL-NifA transcriptional regulatory complex in response to nitrogen, carbon, and redox status. Activation of nif gene expression by the transcriptional activator NifA is inhibited by direct protein-protein interaction with NifL under conditions unfavorable for nitrogen fixation. We have recently shown that the NifL-NifA system responds directly to physiological concentrations of 2-oxoglutarate, resulting in relief of NifA activity from inhibition by NifL under conditions when fixed nitrogen is limiting. The inhibitory activity of NifL is restored under conditions of excess combined nitrogen through the binding of the signal transduction protein Av GlnK to the carboxyl-terminal domain of NifL. The amino-terminal domain of NifA comprises a GAF domain implicated in the regulatory response to NifL. A truncated form of NifA lacking this domain is not responsive to 2-oxoglutarate in the presence of NifL, suggesting that the GAF domain is required for the response to this ligand. Using isothermal titration calorimetry, we demonstrate stoichiometric binding of 2-oxoglutarate to both the isolated GAF domain and the full-length A. vinelandii NifA protein with a dissociation constant of ~60 µM. Limited proteolysis experiments indicate that the binding of 2-oxoglutarate increases the susceptibility of the GAF domain to trypsin digestion and also prevents NifL from protecting these cleavage sites. However, protection by NifL is restored when the non-modified (non-uridylylated) form of Av GlnK is also present. Our results suggest that the binding of 2-oxoglutarate to the GAF domain of NifA may induce a conformational change that prevents inhibition by NifL under conditions when fixed nitrogen is limiting.


Received for publication, February 25, 2003 , and in revised form, May 6, 2003.

* This work was supported by grants from the Biotechnology and Biological Sciences Research Council. 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.

{ddagger} To whom correspondence should be addressed. Tel.: 44-1603-450747; Fax: 44-1603-450778; E-mail: ray.dixon{at}bbsrc.ac.uk.


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


This article has been cited by other articles:


Home page
J. Bacteriol.Home page
T. Iida, T. Waki, K. Nakamura, Y. Mukouzaka, and T. Kudo
The GAF-Like-Domain-Containing Transcriptional Regulator DfdR Is a Sensor Protein for Dibenzofuran and Several Hydrophobic Aromatic Compounds
J. Bacteriol., January 1, 2009; 191(1): 123 - 134.
[Abstract] [Full Text] [PDF]


Home page
MicrobiologyHome page
X. Zou, Y. Zhu, E. L. Pohlmann, J. Li, Y. Zhang, and G. P. Roberts
Identification and functional characterization of NifA variants that are independent of GlnB activation in the photosynthetic bacterium Rhodospirillum rubrum
Microbiology, September 1, 2008; 154(9): 2689 - 2699.
[Abstract] [Full Text] [PDF]


Home page
Eukaryot CellHome page
E. Fernandez and A. Galvan
Nitrate Assimilation in Chlamydomonas
Eukaryot. Cell, April 1, 2008; 7(4): 555 - 559.
[Full Text] [PDF]


Home page
Plant CellHome page
A. Camargo, A. Llamas, R. A. Schnell, J. J. Higuera, D. Gonzalez-Ballester, P. A. Lefebvre, E. Fernandez, and A. Galvan
Nitrate Signaling by the Regulatory Gene NIT2 in Chlamydomonas
PLANT CELL, November 1, 2007; 19(11): 3491 - 3503.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
Y. Qin, S. Su, and S. K. Farrand
Molecular Basis of Transcriptional Antiactivation: TraM DISRUPTS THE TraR-DNA COMPLEX THROUGH STEPWISE INTERACTIONS
J. Biol. Chem., July 6, 2007; 282(27): 19979 - 19991.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
R. Little, I. Martinez-Argudo, S. Perry, and R. Dixon
Role of the H Domain of the Histidine Kinase-like Protein NifL in Signal Transmission
J. Biol. Chem., May 4, 2007; 282(18): 13429 - 13437.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
R. Thummer, O. Klimmek, and R. A. Schmitz
Biochemical Studies of Klebsiella pneumoniae NifL Reduction Using Reconstituted Partial Anaerobic Respiratory Chains of Wolinella succinogenes
J. Biol. Chem., April 27, 2007; 282(17): 12517 - 12526.
[Abstract] [Full Text] [PDF]


Home page
Proc. Natl. Acad. Sci. USAHome page
A. J. Ninfa
Regulation of carbon and nitrogen metabolism: Adding regulation of ion channels and another second messenger to the mix
PNAS, March 13, 2007; 104(11): 4243 - 4244.
[Full Text] [PDF]


Home page
Appl. Environ. Microbiol.Home page
F. E. Rey, E. K. Heiniger, and C. S. Harwood
Redirection of Metabolism for Biological Hydrogen Production
Appl. Envir. Microbiol., March 1, 2007; 73(5): 1665 - 1671.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
S. Bruder, A. Schultz, and J. E. Schultz
Characterization of the Tandem GAF Domain of Human Phosphodiesterase 5 Using a Cyanobacterial Adenylyl Cyclase as a Reporter Enzyme
J. Biol. Chem., July 21, 2006; 281(29): 19969 - 19976.
[Abstract] [Full Text] [PDF]


Home page
J. Bacteriol.Home page
A. Cramer, R. Gerstmeir, S. Schaffer, M. Bott, and B. J. Eikmanns
Identification of RamA, a Novel LuxR-Type Transcriptional Regulator of Genes Involved in Acetate Metabolism of Corynebacterium glutamicum.
J. Bacteriol., April 1, 2006; 188(7): 2554 - 2567.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
M. Gross-Langenhoff, K. Hofbauer, J. Weber, A. Schultz, and J. E. Schultz
cAMP Is a Ligand for the Tandem GAF Domain of Human Phosphodiesterase 10 and cGMP for the Tandem GAF Domain of Phosphodiesterase 11
J. Biol. Chem., February 3, 2006; 281(5): 2841 - 2846.
[Abstract] [Full Text] [PDF]


Home page
Appl. Environ. Microbiol.Home page
R. Mitra, H. K. Das, and A. Dixit
Identification of a Positive Transcription Regulatory Element within the Coding Region of the nifLA Operon in Azotobacter vinelandii
Appl. Envir. Microbiol., July 1, 2005; 71(7): 3716 - 3724.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
R. Zoraghi, E. P. Bessay, J. D. Corbin, and S. H. Francis
Structural and Functional Features in Human PDE5A1 Regulatory Domain That Provide for Allosteric cGMP Binding, Dimerization, and Regulation
J. Biol. Chem., March 25, 2005; 280(12): 12051 - 12063.
[Abstract] [Full Text] [PDF]


Home page
Proc. Natl. Acad. Sci. USAHome page
I. Martinez-Argudo, R. Little, and R. Dixon
A crucial arginine residue is required for a conformational switch in NifL to regulate nitrogen fixation in Azotobacter vinelandii
PNAS, November 16, 2004; 101(46): 16316 - 16321.
[Abstract] [Full Text] [PDF]


Home page
J. Bacteriol.Home page
I. Martinez-Argudo, R. Little, N. Shearer, P. Johnson, and R. Dixon
The NifL-NifA System: a Multidomain Transcriptional Regulatory Complex That Integrates Environmental Signals
J. Bacteriol., February 1, 2004; 186(3): 601 - 610.
[Full Text] [PDF]


Home page
Mol. Pharmacol.Home page
R. Zoraghi, J. D. Corbin, and S. H. Francis
Properties and Functions of GAF Domains in Cyclic Nucleotide Phosphodiesterases and Other Proteins
Mol. Pharmacol., February 1, 2004; 65(2): 267 - 278.
[Full Text] [PDF]




HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement