Protocol

RNA Structure Determination

Volume 1490 of the series Methods in Molecular Biology pp 217-235

Date:

RNA 3D Structure Modeling by Combination of Template-Based Method ModeRNA, Template-Free Folding with SimRNA, and Refinement with QRNAS

  • Pawel PiatkowskiAffiliated withLaboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology 
  • , Joanna M. KasprzakAffiliated withLaboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell BiologyLaboratory of Bioinformatics, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University Email author  
  • , Deepak KumarAffiliated withLaboratory of Bioinformatics, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University 
  • , Marcin MagnusAffiliated withLaboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology
  • , Grzegorz ChojnowskiAffiliated withLaboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology
  • , Janusz M. BujnickiAffiliated withLaboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell BiologyLaboratory of Bioinformatics, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University Email author 

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Abstract

RNA encompasses an essential part of all known forms of life. The functions of many RNA molecules are dependent on their ability to form complex three-dimensional (3D) structures. However, experimental determination of RNA 3D structures is laborious and challenging, and therefore, the majority of known RNAs remain structurally uncharacterized. To address this problem, computational structure prediction methods were developed that either utilize information derived from known structures of other RNA molecules (by way of template-based modeling) or attempt to simulate the physical process of RNA structure formation (by way of template-free modeling). All computational methods suffer from various limitations that make theoretical models less reliable than high-resolution experimentally determined structures. This chapter provides a protocol for computational modeling of RNA 3D structure that overcomes major limitations by combining two complementary approaches: template-based modeling that is capable of predicting global architectures based on similarity to other molecules but often fails to predict local unique features, and template-free modeling that can predict the local folding, but is limited to modeling the structure of relatively small molecules. Here, we combine the use of a template-based method ModeRNA with a template-free method SimRNA. ModeRNA requires a sequence alignment of the target RNA sequence to be modeled with a template of the known structure; it generates a model that predicts the structure of a conserved core and provides a starting point for modeling of variable regions. SimRNA can be used to fold small RNAs (<80 nt) without any additional structural information, and to refold parts of models for larger RNAs that have a correctly modeled core. ModeRNA can be either downloaded, compiled and run locally or run through a web interface at http://​genesilico.​pl/​modernaserver/​. SimRNA is currently available to download for local use as a precompiled software package at http://​genesilico.​pl/​software/​stand-alone/​simrna and as a web server at http://​genesilico.​pl/​SimRNAweb. For model optimization we use QRNAS, available at http://​genesilico.​pl/​qrnas.

Key words

RNA structure Comparative modeling Homology modeling Free modeling De novo modeling Monte Carlo simulation Statistical potential