Mechanism of MicroRNA-Target Interaction: Molecular Dynamics Simulations and Thermodynamics Analysis

MicroRNAs (miRNAs) are endogenously produced ∼21-nt riboregulators that associate with Argonaute (Ago) proteins to direct mRNA cleavage or repress the translation of complementary RNAs. Capturing the molecular mechanisms of miRNA interacting with its target will not only reinforce the understanding of underlying RNA interference but also fuel the design of more effective small-interfering RNA strands. To address this, in the present work the RNA-bound (Ago-miRNA, Ago-miRNA-target) and RNA-free Ago forms were analyzed by performing both molecular dynamics simulations and thermodynamic analysis. Based on the principal component analysis results of the simulation trajectories as well as the correlation analysis in fluctuations of residues, we discover that: 1) three important (PAZ, Mid and PIWI) domains exist in Argonaute which define the global dynamics of the protein; 2) the interdomain correlated movements are so crucial for the interaction of Ago-RNAs that they not only facilitate the relaxation of the interactions between residues surrounding the RNA binding channel but also induce certain conformational changes; and 3) it is just these conformational changes that expand the cavity of the active site and open putative pathways for both the substrate uptake and product release. In addition, by thermodynamic analysis we also discover that for both the guide RNA 5′-end recognition and the facilitated site-specific cleavage of the target, the presence of two metal ions (of Mg2+) plays a predominant role, and this conclusion is consistent with the observed enzyme catalytic cleavage activity in the ternary complex (Ago-miRNA-mRNA). Our results find that it is the set of arginine amino acids concentrated in the nucleotide-binding channel in Ago, instead of the conventionally-deemed seed base-paring, that makes greater contributions in stabilizing the binding of the nucleic acids to Ago.Author Summary: One of the biggest surprises at the beginning of the ‘post-genome era’ was the discovery of numerous genes encoding microRNAs. The number of microRNA genes is estimated to be nearly 1% of that of protein-coding genes, which were found in genomes of such diverse organisms as Caenorhabditis elegans, Drosophila melanogaster, Arabidopsis thaliana, and Homo sapiens. Their products, tiny RNAs (miRNAs and siRNAs), are thought to bind to Argonaute (Ago) proteins and form effector complexes to direct mRNA cleavage or repress translation of complementary RNAs, during development, organogenesis, and very likely during many other processes. The cellular interactions between the miRNAs and their target RNAs associating with Ago are only beginning to be revealed, and details of this interaction mechanism at molecular level are still poorly understood. In this article we propose the possible mechanisms of miRNA-target interaction with special emphasis on their structural dynamic and thermodynamic aspects. The results of our model suggest the chemical and physical factors and effects that may be responsible for the miRNA-Ago assembly and miRNA-target recognition.