Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 25 March 2005:
Vol. 307. no. 5717, pp. 1972 - 1975
DOI: 10.1126/science.1105932
Prev | Table of Contents | Next

Reports

Structural Insights into the Activity of Enhancer-Binding Proteins

Mathieu Rappas,1,2 Jorg Schumacher,1 Fabienne Beuron,1,2 Hajime Niwa,1,2 Patricia Bordes,1 Sivaramesh Wigneshweraraj,1 Catherine A. Keetch,3 Carol V. Robinson,3 Martin Buck,1 Xiaodong Zhang1,2*

Activators of bacterial {sigma}54–RNA polymerase holoenzyme are mechanochemical proteins that use adenosine triphosphate (ATP) hydrolysis to activate transcription. We have determined by cryogenic electron microscopy (cryo-EM) a 20 angstrom resolution structure of an activator, phage shock protein F [PspF(1-275)], which is bound to an ATP transition state analog in complex with its basal factor, {sigma}54. By fitting the crystal structure of PspF(1-275) at 1.75 angstroms into the EM map, we identified two loops involved in binding {sigma}54. Comparing enhancer-binding structures in different nucleotide states and mutational analysis led us to propose nucleotide-dependent conformational changes that free the loops for association with {sigma}54.

1 Department of Biological Sciences, Imperial College London, London, SW7 2AZ, UK.
2 Centre for Structural Biology, Imperial College London, London, SW7 2AZ, UK.
3 Department of Chemistry, Cambridge University, Lensfield Road, Cambridge, CB2 1EW, UK.

* To whom correspondence should be addressed. E-mail: xiaodong.zhang{at}imperial.ac.uk

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
The role of the conserved phenylalanine in the {sigma}54-interacting GAFTGA motif of bacterial enhancer binding proteins.
N. Zhang, N. Joly, P. C. Burrows, M. Jovanovic, S. R. Wigneshweraraj, and M. Buck (2009)
Nucleic Acids Res. 37, 5981-5992
   Abstract »    Full Text »    PDF »
Comparative analysis of activator-E{sigma}54 complexes formed with nucleotide-metal fluoride analogues.
P. C. Burrows, N. Joly, B. T. Nixon, and M. Buck (2009)
Nucleic Acids Res. 37, 5138-5150
   Abstract »    Full Text »    PDF »
Rapid detection, classification and accurate alignment of up to a million or more related protein sequences.
A. F. Neuwald (2009)
Bioinformatics 25, 1869-1875
   Abstract »    Full Text »    PDF »
Construction and functional analyses of a comprehensive {sigma}54 site-directed mutant library using alanine-cysteine mutagenesis.
Y. Xiao, S. R. Wigneshweraraj, R. Weinzierl, Y.-P. Wang, and M. Buck (2009)
Nucleic Acids Res. 37, 4482-4497
   Abstract »    Full Text »    PDF »
Identification of the binding site of the {sigma}54 hetero-oligomeric FleQ/FleT activator in the flagellar promoters of Rhodobacter sphaeroides.
J. Pena-Sanchez, S. Poggio, U. Flores-Perez, A. Osorio, C. Domenzain, G. Dreyfus, and L. Camarena (2009)
Microbiology 155, 1669-1679
   Abstract »    Full Text »    PDF »
The bacterial type VI secretion machine: yet another player for protein transport across membranes.
A. Filloux, A. Hachani, and S. Bleves (2008)
Microbiology 154, 1570-1583
   Abstract »    Full Text »    PDF »
An Intramolecular Route for Coupling ATPase Activity in AAA+ Proteins for Transcription Activation.
N. Joly, P. C. Burrows, and M. Buck (2008)
J. Biol. Chem. 283, 13725-13735
   Abstract »    Full Text »    PDF »
Replication Origin Recognition and Deformation by a Heterodimeric Archaeal Orc1 Complex.
E. L. C. Dueber, J. E. Corn, S. D. Bell, and J. M. Berger (2007)
Science 317, 1210-1213
   Abstract »    Full Text »    PDF »
Sensor I Threonine of the AAA+ ATPase Transcriptional Activator PspF Is Involved in Coupling Nucleotide Triphosphate Hydrolysis to the Restructuring of {sigma}54-RNA Polymerase.
J. Schumacher, N. Joly, M. Rappas, D. Bradley, S. R. Wigneshweraraj, X. Zhang, and M. Buck (2007)
J. Biol. Chem. 282, 9825-9833
   Abstract »    Full Text »    PDF »
Use of plasmon coupling to reveal the dynamics of DNA bending and cleavage by single EcoRV restriction enzymes.
B. M. Reinhard, S. Sheikholeslami, A. Mastroianni, A. P. Alivisatos, and J. Liphardt (2007)
PNAS 104, 2667-2672
   Abstract »    Full Text »    PDF »
A Role for the Conserved GAFTGA Motif of AAA+ Transcription Activators in Sensing Promoter DNA Conformation.
A. E. Dago, S. R. Wigneshweraraj, M. Buck, and E. Morett (2007)
J. Biol. Chem. 282, 1087-1097
   Abstract »    Full Text »    PDF »
A structural model reveals energy transduction in dynein.
A. W. R. Serohijos, Y. Chen, F. Ding, T. C. Elston, and N. V. Dokholyan (2006)
PNAS 103, 18540-18545
   Abstract »    Full Text »    PDF »
Heterogeneous Nucleotide Occupancy Stimulates Functionality of Phage Shock Protein F, an AAA+ Transcriptional Activator.
N. Joly, J. Schumacher, and M. Buck (2006)
J. Biol. Chem. 281, 34997-35007
   Abstract »    Full Text »    PDF »
Structural basis of the Methanothermobacter thermautotrophicus MCM helicase activity.
A. Costa, T. Pape, M. van Heel, P. Brick, A. Patwardhan, and S. Onesti (2006)
Nucleic Acids Res. 34, 5829-5838
   Abstract »    Full Text »    PDF »
Mapping ATP-dependent Activation at a {sigma}54 Promoter.
R. N. Leach, C. Gell, S. Wigneshweraraj, M. Buck, A. Smith, and P. G. Stockley (2006)
J. Biol. Chem. 281, 33717-33726
   Abstract »    Full Text »    PDF »
Transcriptional Specificity of RpoN1 and RpoN2 Involves Differential Recognition of the Promoter Sequences and Specific Interaction with the Cognate Activator Proteins.
S. Poggio, A. Osorio, G. Dreyfus, and L. Camarena (2006)
J. Biol. Chem. 281, 27205-27215
   Abstract »    Full Text »    PDF »
The structural basis for regulated assembly and function of the transcriptional activator NtrC.
S. De Carlo, B. Chen, T. R. Hoover, E. Kondrashkina, E. Nogales, and B. T. Nixon (2006)
Genes & Dev. 20, 1485-1495
   Abstract »    Full Text »    PDF »
The Upstream-activating Sequences of the {sigma}54 Promoter Pu of Pseudomonas putida Filter Transcription Readthrough from Upstream Genes.
F. Velazquez, S. Fernandez, and V. de Lorenzo (2006)
J. Biol. Chem. 281, 11940-11948
   Abstract »    Full Text »    PDF »
The C-terminal RpoN Domain of {sigma}54 Forms an Unpredicted Helix-Turn-Helix Motif Similar to Domains of {sigma}70.
M. Doucleff, L. T. Malak, J. G. Pelton, and D. E. Wemmer (2005)
J. Biol. Chem. 280, 41530-41536
   Abstract »    Full Text »    PDF »
Stable DNA Opening within Open Promoter Complexes Is Mediated by the RNA Polymerase {beta}'-Jaw Domain.
Siva. R. Wigneshweraraj, P. C. Burrows, K. Severinov, and M. Buck (2005)
J. Biol. Chem. 280, 36176-36184
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)