title: Biochemical Methods To Investigate lncRNA and the Influence of lncRNA:Protein Complexes on Chromatin.
abstract: Long noncoding RNAs (lncRNAs), defined as nontranslated transcripts greater than 200 nucleotides in length, are often differentially expressed throughout developmental stages, tissue types, and disease states. The identification, visualization, and suppression/overexpression of these sequences have revealed impacts on a wide range of biological processes, including epigenetic regulation. Biochemical investigations on select systems have revealed striking insight into the biological roles of lncRNAs and lncRNA:protein complexes, which in turn prompt even more unanswered questions. To begin, multiple protein- and RNA-centric technologies have been employed to isolate lncRNA:protein and lncRNA:chromatin complexes. LncRNA interactions with the multi-subunit protein complex PRC2, which acts as a transcriptional silencer, represent some of the few cases where the binding affinity, selectivity, and activity of a lncRNA:protein complex have been investigated. At the same time, recent reports of full-length lncRNA secondary structures suggest the formation of complex structures with multiple independent folding domains and pave the way for more detailed structural investigations and predictions of lncRNA three-dimensional structure. This review will provide an overview of the methods and progress made to date as well as highlight new methods that promise to further inform the molecular recognition, specificity, and function of lncRNAs.
authors: Department of Biochemistry, Duke University Medical Center , Durham, North Carolina 27710, United States.
authors: Emily J
authors: EJ
authors: McFadden
authors: Department of Biochemistry, Duke University Medical Center , Durham, North Carolina 27710, United States.|Department of Chemistry, Duke University , 124 Science Drive, Durham, North Carolina 27708, United States.
authors: Amanda E
authors: AE
authors: Hargrove
mesh_terms: D000818:Animals; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology*; D006801:Humans; D063151:Polycomb Repressive Complex 2 / Q000378:metabolism*; D062085:RNA, Long Noncoding / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*
chemical_list: D002843:Chromatin; D062085:RNA, Long Noncoding; D012261:Ribonucleoproteins; D063151:Polycomb Repressive Complex 2