A Caltech Library Service

RNA–protein recognition: Single-residue ultrafast dynamical control of structural specificity and function

Xia, Tianbing and Wan, Chaozhi and Roberts, Richard W. and Zewail, Ahmed H. (2005) RNA–protein recognition: Single-residue ultrafast dynamical control of structural specificity and function. Proceedings of the National Academy of Sciences of the United States of America, 102 (37). pp. 13013-13018. ISSN 0027-8424. PMCID PMC1201610.

PDF - Published Version
See Usage Policy.

[img] PDF (Fig. 7. Initial signal intensity measured in fluorescence upconversion (Upper) and transient absorption (Lower) collected in a short time window ...) - Supplemental Material
See Usage Policy.

[img] PDF (Fig. 8. Effects of substitution of Glu-13 by proline on the dynamics measured in both transient absorption (Top) and fluorescence upconversion experiments (Middle) and on transcription antitermination activity level in vivo (Bottom) ...) - Supplemental Material
See Usage Policy.


Use this Persistent URL to link to this item:


The transcription antiterminator IN protein from bacteriophage x uses its arginine-rich motif to specifically bind a stem-loop RNA hairpin (boxB) as a bent alpha-helix. A single stacking interaction between a tryptophan (Trp-18) and an adenosine (A7) in the RNA loop is robust and necessary for antitermination activity in vivo. Previously, femtosecond fluorescence up-conversion experiments from this laboratory indicated that the N/boxB complex exists in a dynamical two-state equilibrium between stacked and unstacked conformations and that the extent of stacking depends on the identity of peptide residues 14 and 15. In the present work, we have combined transient absorption and fluorescence up-conversion to determine the nature of interactions responsible for this sequence-dependent behavior. Analysis of mutant complexes supports the idea that the beta-carbon of residue 14 enforces the stacked geometry by hydrophobic interaction with the ribose of A7, whereas a positive charge at this residue plays only a secondary role. A positive charge at position 15 substantially disfavors the stacked state but retains much of the binding energy. Remarkably, in vivo antitermination experiments show strong correlation with our femtosecond dynamics, demonstrating how conformational interplay can control the activity of a macromolecular machine.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle Information
Roberts, Richard W.0000-0002-8587-5097
Additional Information:© 2005 by the National Academy of Sciences. Contributed by Ahmed H. Zewail, August 2, 2005. Published online before print August 29, 2005, 10.1073/pnas.0506181102. We thank Prof. Naomi Franklin (University of Utah, Salt Lake City) for the two-plasmid N expressor-β-gal reporter constructs; Prof. Douglas Turner for helpful comments, Dr. Adam Frankel (now at University of British Columbia, Vancouver) for help with the transcription antitermination assay, Dr. Suzanna Horvath for purification of some peptides used in this work, and Ms. Nancy Guillen for help with measurements of binding constants. This work was supported by the National Science Foundation through the Laboratory of Molecular Sciences, making it possible to integrate expertise in chemical physics and molecular biology.
Funding AgencyGrant Number
Issue or Number:37
PubMed Central ID:PMC1201610
Record Number:CaltechAUTHORS:XIApnas05
Persistent URL:
Official Citation:Tianbing Xia, Chaozhi Wan, Richard W. Roberts, and Ahmed H. Zewail RNA–protein recognition: Single-residue ultrafast dynamical control of structural specificity and function PNAS 2005 102 (37) 13013-13018; published ahead of print August 29, 2005, doi:10.1073/pnas.0506181102
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:2134
Deposited By: Tony Diaz
Deposited On:10 Mar 2006
Last Modified:09 Mar 2020 13:19

Repository Staff Only: item control page