![Show Menu](styles/mobile-menu.png)
![Page Background](./../common/page-substrates/page0101.jpg)
Conformational Ensembles from Experimental Data
and Computer Simulations
Poster Abstracts
97
62-POS
Board 22
A Simulation-based Approach to the Dynamical Basis of Hfq-RNA Interactions
Charles McAnany
, Sooraj Achar, Kimberly Stanek, Cameron Mura.
University of Virginia, Charlottesville, VA, USA.
The bacterial Sm protein, known as Hfq, acts as a generic RNA chaperone that facilitates
interactions between two RNA strands, typically a noncoding small RNA (sRNA) and a
regulatory target (e.g., an mRNA). Many sRNAs play key roles in post-transcriptional
regulation, including protein translational control and RNA decay pathways. While a decade of
crystal structures have provided static snapshots of Hfq and Hfq-RNA complexes, and
biochemical studies have supplied valuable information about Hfq function, the physicochemical
behavior of RNA and Hfq, as they interact across their molecular surfaces, remains unexplored.
Hfq self-assembles into hexameric rings, with two distinct RNA-binding regions. One side of the
ring (the
distal
face) binds U-rich RNA, while the other (
proximal
) face binds A-rich RNAs. Our
recent crystal structure of an
Aquifex aeolicus
Hfq reveals, in addition to the proximal and distal
sites, a conserved
lateral
RNA site on the periphery of the ring. This lateral site binds U-rich
RNA with lower affinity than the proximal site. To see how RNA interacts with Hfq, we are
pursuing an extensive suite of µsec-scale MD simulations. By using steered MD to drive two
nucleotides of RNA toward the lateral site, our simulations start with a physically plausible,
partially-bound state. We then simulate the unconstrained system to examine RNA interactions
with the neighboring protein surface, guided by specific questions such as: Can RNA
simultaneously bind both the lateral and distal (or lateral and proximal) sites? How stable and
persistent (thermodynamically and kinetically) are RNA interactions with the lateral site? These
simulations will illuminate, in molecular detail, the fundamental mechanism of Hfq-mediated
RNA annealing.