PeproTech, Our Business is Cytokines! Sign up for PNAS Online eTocs
Link: Info for AuthorsLink: Editorial BoardLink: AboutLink: SubscribeLink: AdvertiseLink: ContactLink: Sitemap Link: PNAS Home
Proceedings of the National Academy of Sciences
Link: Current Issue "" Link: Archives "" Link: Online Submission ""  Link: Advanced Search

Published online on May 27, 2003, 10.1073/pnas.1037608100
PNAS | June 10, 2003 | vol. 100 | no. 12 | 7099-7104


This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supporting Information
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a colleague
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My File Cabinet
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via ISI Web of Science (8)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lopian, L.
Right arrow Articles by Amster-Choder, O.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lopian, L.
Right arrow Articles by Amster-Choder, O.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg  
What's this?

 Previous Article  | Table of Contents |  Next Article 

Cell Biology
The BglF sensor recruits the BglG transcription regulator to the membrane and releases it on stimulation

Livnat Lopian *, Anat Nussbaum-Shochat *, Kathryn O'Day-Kerstein {dagger}, Andrew Wright {dagger}, and Orna Amster-Choder * {ddagger}

*Department of Molecular Biology, The Hebrew University–Hadassah Medical School, P.O. Box 12272, Jerusalem 91120, Israel; and{dagger}Department of Molecular Biology and Microbiology, Tufts University Health Sciences Campus, Boston, MA 02111

Edited by Charles Yanofsky, Stanford University, Stanford, CA and approved March 17, 2003 (received for review December 15, 2002)

The Escherichia coli BglF protein is a sugar-sensor that controls the activity of the transcriptional antiterminator BglG by reversibly phosphorylating it, depending on {beta}-glucoside availability. BglF is a membrane-bound protein, whereas BglG is a soluble protein, and they are both present in the cell in minute amounts. How do BglF and BglG find each other to initiate signal transduction efficiently? Using bacterial two-hybrid systems and the Far-Western technique, we demonstrated unequivocally that BglG binds to BglF and to its active site-containing domain in vivo and in vitro. Measurements by surface plasmon resonance corroborated that the affinity between these proteins is high enough to enable their stable binding. To visualize the subcellular localization of BglG, we used fluorescence microscopy. In cells lacking BglF, the BglG-GFP fusion protein was evenly distributed throughout the cytoplasm. In contrast, in cells producing BglF, BglG-GFP was localized to the membrane. On addition of {beta}-glucoside, BglG-GFP was released from the membrane, becoming evenly distributed throughout the cell. Using mutant proteins and genetic backgrounds that impede phosphorylation of the Bgl proteins, we demonstrated that BglG-BglF binding and recruitment of BglG to the membrane sensor requires phosphorylation but does not depend on the individual phosphorylation sites of the Bgl proteins. We suggest a mechanism for rapid response to environmental changes by preassembly of signaling complexes, which contain transcription regulators recruited by their cognate sensors-kinases, under nonstimulating conditions, and release of the regulators to the cytoplasm on stimulation. This mechanism might be applicable to signaling cascades in prokaryotes and eukaryotes.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: PTS, phosphotransferase system; SPR, surface plasmon resonance; MBP, maltose-binding protein; LexADBD, LexA repressor DNA-binding domain.

{ddagger} To whom correspondence should be addressed. E-mail: amster{at}cc.huji.ac.il.


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


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


Home page
 J. Bacteriol.Home page
G. Monderer-Rothkoff and O. Amster-Choder
Genetic Dissection of the Divergent Activities of the Multifunctional Membrane Sensor BglF
J. Bacteriol., December 1, 2007; 189(23): 8601 - 8615.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
Z. Pang, R. Chen, D. Manna, and N. P. Higgins
A Gyrase Mutant with Low Activity Disrupts Supercoiling at the Replication Terminus
J. Bacteriol., November 15, 2005; 187(22): 7773 - 7783.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Graille, C.-Z. Zhou, V. Receveur-Brechot, B. Collinet, N. Declerck, and H. van Tilbeurgh
Activation of the LicT Transcriptional Antiterminator Involves a Domain Swing/Lock Mechanism Provoking Massive Structural Changes
J. Biol. Chem., April 15, 2005; 280(15): 14780 - 14789.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
L. Fux, A. Nussbaum-Shochat, L. Lopian, and O. Amster-Choder
Modulation of Monomer Conformation of the BglG Transcriptional Antiterminator from Escherichia coli
J. Bacteriol., October 15, 2004; 186(20): 6775 - 6781.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
A. S. Johnson, S. van Horck, and P. J. Lewis
Dynamic localization of membrane proteins in Bacillus subtilis
Microbiology, September 1, 2004; 150(9): 2815 - 2824.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
N. Joly, A. Bohm, W. Boos, and E. Richet
MalK, the ATP-binding Cassette Component of the Escherichia coli Maltodextrin Transporter, Inhibits the Transcriptional Activator MalT by Antagonizing Inducer Binding
J. Biol. Chem., August 6, 2004; 279(32): 33123 - 33130.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Deng, R. A. Stein, and N. P. Higgins
Transcription-induced barriers to supercoil diffusion in the Salmonella typhimurium chromosome
PNAS, March 9, 2004; 101(10): 3398 - 3403.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. Fux, A. Nussbaum-Shochat, and O. Amster-Choder
A Fraction of the BglG Transcriptional Antiterminator from Escherichia coli Exists as a Compact Monomer
J. Biol. Chem., December 19, 2003; 278(51): 50978 - 50984.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. Fux, A. Nussbaum-Shochat, and O. Amster-Choder
Interactions between the PTS Regulation domains of the BglG Transcriptional Antiterminator from Escherichia coli
J. Biol. Chem., November 21, 2003; 278(47): 46203 - 46209.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
B. Gorke
Regulation of the Escherichia coli Antiterminator Protein BglG by Phosphorylation at Multiple Sites and Evidence for Transfer of Phosphoryl Groups between Monomers
J. Biol. Chem., November 21, 2003; 278(47): 46219 - 46229.
[Abstract] [Full Text] [PDF]