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      Whole genome experimental maps of DNA G-quadruplexes in multiple species

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          Abstract

          Genomic maps of DNA G-quadruplexes (G4s) can help elucidate the roles that these secondary structures play in various organisms. Herein, we employ an improved version of a G-quadruplex sequencing method (G4-seq) to generate whole genome G4 maps for 12 species that include widely studied model organisms and also pathogens of clinical relevance. We identify G4 structures that form under physiological K + conditions and also G4s that are stabilized by the G4-targeting small molecule pyridostatin (PDS). We discuss the various structural features of the experimentally observed G-quadruplexes (OQs), highlighting differences in their prevalence and enrichment across species. Our study describes diversity in sequence composition and genomic location for the OQs in the different species and reveals that the enrichment of OQs in gene promoters is particular to mammals such as mouse and human, among the species studied. The multi-species maps have been made publicly available as a resource to the research community. The maps can serve as blueprints for biological experiments in those model organisms, where G4 structures may play a role.

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          Stability and kinetics of G-quadruplex structures

          In this review, we give an overview of recent literature on the structure and stability of unimolecular G-rich quadruplex structures that are relevant to drug design and for in vivo function. The unifying theme in this review is energetics. The thermodynamic stability of quadruplexes has not been studied in the same detail as DNA and RNA duplexes, and there are important differences in the balance of forces between these classes of folded oligonucleotides. We provide an overview of the principles of stability and where available the experimental data that report on these principles. Significant gaps in the literature have been identified, that should be filled by a systematic study of well-defined quadruplexes not only to provide the basic understanding of stability both for design purposes, but also as it relates to in vivo occurrence of quadruplexes. Techniques that are commonly applied to the determination of the structure, stability and folding are discussed in terms of information content and limitations. Quadruplex structures fold and unfold comparatively slowly, and DNA unwinding events associated with transcription and replication may be operating far from equilibrium. The kinetics of formation and resolution of quadruplexes, and methodologies are discussed in the context of stability and their possible biological occurrence.
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            Gene function correlates with potential for G4 DNA formation in the human genome

            G-rich genomic regions can form G4 DNA upon transcription or replication. We have quantified the potential for G4 DNA formation (G4P) of the 16 654 genes in the human RefSeq database, and then correlated gene function with G4P. We have found that very low and very high G4P correlates with specific functional classes of genes. Notably, tumor suppressor genes have very low G4P and proto-oncogenes have very high G4P. G4P of these genes is evenly distributed between exons and introns, and it does not reflect enrichment for CpG islands or local chromosomal environment. These results show that genomic structure undergoes selection based on gene function. Selection based on G4P could promote genomic stability (or instability) of specific classes of genes; or reflect mechanisms for global regulation of gene expression.
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              rG4-seq reveals widespread formation of G-quadruplex structures in the human transcriptome.

              We introduce RNA G-quadruplex sequencing (rG4-seq), a transcriptome-wide RNA G-quadruplex (rG4) profiling method that couples rG4-mediated reverse transcriptase stalling with next-generation sequencing. Using rG4-seq on polyadenylated-enriched HeLa RNA, we generated a global in vitro map of thousands of canonical and noncanonical rG4 structures. We characterize rG4 formation relative to cytosine content and alternative RNA structure stability, uncover rG4-dependent differences in RNA folding and show evolutionarily conserved enrichment in transcripts mediating RNA processing and stability.
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                Author and article information

                Journal
                Nucleic Acids Res
                Nucleic Acids Res
                nar
                Nucleic Acids Research
                Oxford University Press
                0305-1048
                1362-4962
                07 May 2019
                20 March 2019
                20 March 2019
                : 47
                : 8
                : 3862-3874
                Affiliations
                [1 ]Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Cambridge CB2 0RE, UK
                [2 ]Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK
                [3 ]Illumina Cambridge Ltd., Chesterford Research Park, Little Chesterford, Saffron Walden, Essex CB10 1XL, UK
                [4 ]School of Clinical Medicine, University of Cambridge, Cambridge CB2 0SP, UK
                Author notes
                To whom correspondence should be addressed. Tel: +44 1223 336347; Email: sb10031@ 123456cam.ac.uk
                Present address: Marco Di Antonio, Department of Chemistry, Molecular Science Research Hub, Imperial College London, London W12 0BZ, UK
                Present address: Aleksandr B Sahakyan, MRC WIMM Centre for Computational Biology, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UK
                Author information
                http://orcid.org/0000-0002-8343-3594
                http://orcid.org/0000-0002-0281-5815
                Article
                gkz179
                10.1093/nar/gkz179
                6486626
                30892612
                be060654-c7af-4b09-a9a4-78495d5947a7
                © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 08 March 2019
                : 04 March 2019
                : 29 November 2018
                Page count
                Pages: 13
                Funding
                Funded by: Cancer Research UK 10.13039/501100000289
                Award ID: C9681/A18618
                Funded by: European Research Council 10.13039/501100000781
                Award ID: 339778
                Funded by: Wellcome Trust 10.13039/100004440
                Award ID: 209441/z/17/z
                Funded by: Cancer Research UK 10.13039/501100000289
                Award ID: C14303/A17197
                Funded by: Biotechnology and Biological Sciences Research Council 10.13039/501100000268
                Award ID: BB/I015477/1
                Categories
                Data Resources and Analyses

                Genetics
                Genetics

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