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      A comprehensive database of high-throughput sequencing-based RNA secondary structure probing data (Structure Surfer)

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          Abstract

          Background

          RNA molecules fold into complex three-dimensional shapes, guided by the pattern of hydrogen bonding between nucleotides. This pattern of base pairing, known as RNA secondary structure, is critical to their cellular function. Recently several diverse methods have been developed to assay RNA secondary structure on a transcriptome-wide scale using high-throughput sequencing. Each approach has its own strengths and caveats, however there is no widely available tool for visualizing and comparing the results from these varied methods.

          Methods

          To address this, we have developed Structure Surfer, a database and visualization tool for inspecting RNA secondary structure in six transcriptome-wide data sets from human and mouse ( http://tesla.pcbi.upenn.edu/strucuturesurfer/). The data sets were generated using four different high-throughput sequencing based methods. Each one was analyzed with a scoring pipeline specific to its experimental design. Users of Structure Surfer have the ability to query individual loci as well as detect trends across multiple sites.

          Results

          Here, we describe the included data sets and their differences. We illustrate the database’s function by examining known structural elements and we explore example use cases in which combined data is used to detect structural trends.

          Conclusions

          In total, Structure Surfer provides an easy-to-use database and visualization interface for allowing users to interrogate the currently available transcriptome-wide RNA secondary structure information for mammals.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12859-016-1071-0) contains supplementary material, which is available to authorized users.

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          Most cited references6

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          A fast-acting reagent for accurate analysis of RNA secondary and tertiary structure by SHAPE chemistry.

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            Direct chemical method for sequencing RNA.

            Four different base-specific chemical reactions generate a means of directly sequencing RNA terminally labeled with 32P. After a partial, specific modification of each kind of RNA base, an amine-catalyzed strand scission generates labeled fragments whose lengths determine the position of each nucleotide in the sequence. Dimethyl sulfate modifies guanosine. Diethyl pyrocarbonate attacks primarily adenosine. Hydrazine attacks uridine and cytidine, but salt suppresses the reaction with uridine. In all cases, aniline induces a subsequent strand scission. The electrophoretic fractionation of the labeled fragments on a polyacrylamide gel, followed by autoradiography, determines the RNA sequence. RNA labeled at the 3' end yields clean cleavage patterns for each purine and pyrimidine and allows a determination of the entire RNA sequence out to 100-200 bases from the labeled terminus.
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              Snapshots of pre-rRNA structural flexibility reveal eukaryotic 40S assembly dynamics at nucleotide resolution

              Ribosome assembly in eukaryotes involves the activity of hundreds of assembly factors that direct the hierarchical assembly of ribosomal proteins and numerous ribosomal RNA folding steps. However, detailed insights into the function of assembly factors and ribosomal RNA folding events are lacking. To address this, we have developed ChemModSeq, a method that combines structure probing, high-throughput sequencing and statistical modeling, to quantitatively measure RNA structural rearrangements during the assembly of macromolecular complexes. By applying ChemModSeq to purified 40S assembly intermediates we obtained nucleotide-resolution maps of ribosomal RNA flexibility revealing structurally distinct assembly intermediates and mechanistic insights into assembly dynamics not readily observed in cryo-electron microscopy reconstructions. We show that RNA restructuring events coincide with the release of assembly factors and predict that completion of the head domain is required before the Rio1 kinase enters the assembly pathway. Collectively, our results suggest that 40S assembly factors regulate the timely incorporation of ribosomal proteins by delaying specific folding steps in the 3′ major domain of the 20S pre-ribosomal RNA.
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                Author and article information

                Contributors
                (215) 746-4398 , (215) 898-8780 , bdgregor@sas.upenn.edu
                Journal
                BMC Bioinformatics
                BMC Bioinformatics
                BMC Bioinformatics
                BioMed Central (London )
                1471-2105
                17 May 2016
                17 May 2016
                2016
                : 17
                : 215
                Affiliations
                [ ]Department of Biology, University of Pennsylvania, 433 S. University Ave., Philadelphia, PA 19104 USA
                [ ]Genomics and Computational Biology Graduate Group, Philadelphia, USA
                [ ]Cell and Molecular Biology Graduate Group, Philadelphia, USA
                [ ]Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104 USA
                [ ]Department of Computer Engineering, Antalya International University, Antalya, Turkey
                [ ]Institute on Aging, Baltimore, USA
                [ ]Penn Center for Bioinformatics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104 USA
                Article
                1071
                10.1186/s12859-016-1071-0
                4869249
                27188311
                704718fa-d3f3-4c25-b9e1-de2cc63b084c
                © Berkowitz et al. 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 4 March 2016
                : 4 May 2016
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: MCB-1053846, MCB-1243947, and IOS-1444490
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000057, National Institute of General Medical Sciences;
                Award ID: R01-GM099962 and 5T32GM008216-26
                Award Recipient :
                Funded by: Marie Curie CIG Grant
                Award ID: 631986
                Award Recipient :
                Funded by: National Institute of Health
                Award ID: U24-AG041689
                Award Recipient :
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                © The Author(s) 2016

                Bioinformatics & Computational biology
                Bioinformatics & Computational biology

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