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Assessing Intermolecular RNA:RNA Interactions Within a Ribonucleoprotein Complex Using Heavy Metal Cleavage Mapping

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RNA-RNA Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1240))

Abstract

Heavy metal cleavage mapping analysis of both assembling and fully mature ribonucleoprotein (RNP) complexes are informative techniques for assessing the intermolecular base pairing between small non-coding RNAs and their interacting target RNAs. Lead cleavage of the RNA in partially or fully assembled RNPs in the absence or presence of the interacting RNA can determine both the accessibility of the base pairing sequence within the RNP itself as well as its interaction with the target RNA. In this chapter, we detail how this technique was used to map the intermolecular RNA:RNA base pairing of a box C/D RNA with its target RNA within the assembling archaeal RNP complex.

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References

  1. Valadkhan S (2010) Role of the snRNAs in spliceosomal active site. RNA Biol 7:345–353

    Article  CAS  PubMed  Google Scholar 

  2. Bartel D (2004) MiRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297

    Article  CAS  PubMed  Google Scholar 

  3. Terns M, Terns R (2002) Small nucleolar RNAs: versatile trans-acting molecules of ancient evolutionary origin. Gene Expr 10:17–39

    CAS  PubMed  Google Scholar 

  4. Gagnon KT, Zhang X, Agris PF, Maxwell ES (2006) Assembly of the archaeal box C/D sRNP can occur via alternative pathways and requires temperature-facilitated sRNA remodeling. J Mol Biol 362(5):1025–1042

    Article  CAS  PubMed  Google Scholar 

  5. Gagnon KT, Maxwell ES (2011) Electrophoretic mobility shift assay for characterizing rna-protein interaction. In: Nielsen H (ed) Methods in molecular biology, vol 703. Humana, New York, pp 275–291

    Google Scholar 

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Correspondence to Keith T. Gagnon Ph.D. or E. Stuart Maxwell Ph.D. .

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© 2015 Springer Science+Business Media New York

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Gagnon, K.T., Maxwell, E.S. (2015). Assessing Intermolecular RNA:RNA Interactions Within a Ribonucleoprotein Complex Using Heavy Metal Cleavage Mapping. In: Schmidt, F. (eds) RNA-RNA Interactions. Methods in Molecular Biology, vol 1240. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1896-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-1896-6_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1895-9

  • Online ISBN: 978-1-4939-1896-6

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