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In Vitro Analysis of Ribonucleoprotein Complex Remodeling and Disassembly

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RNA-Protein Complexes and Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1421))

Abstract

Ribonucleoprotein (RNP) complexes play essential roles in gene expression. Their assembly and disassembly control the fate of mRNA molecules. Here, we describe a method that examines the remodeling and disassembly of RNPs. One unique aspect of this method is that the RNA-binding proteins (RBPs) of interest are produced in HeLa cells with or without the desired modification and the RNP is assembled in cellular extracts with synthetic RNA oligonucleotides. We use this method to investigate how ubiquitination of an RBP affects its ability to bind its RNA target.

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References

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Acknowledgements

This work was supported by an NIH grant (NS-049103) and a DOD grant (NF060083) to Hua Lou. Hua-Lin Zhou was supported by post-doctoral fellowships from the American Heart Association (0725346B and 09POST2250749).

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Authors and Affiliations

  1. Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH, USA

    Hua-Lin Zhou & Hua Lou

Authors
  1. Hua-Lin Zhou
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  2. Hua Lou
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Corresponding author

Correspondence to Hua-Lin Zhou .

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Editors and Affiliations

  1. Beckman res.Ins.Dept.of Mol. & Cell Bio., Irell&Manella.Grad.Schl.Biolog.Sci., Duarte, California, USA

    Ren-Jang Lin

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Zhou, HL., Lou, H. (2016). In Vitro Analysis of Ribonucleoprotein Complex Remodeling and Disassembly. In: Lin, RJ. (eds) RNA-Protein Complexes and Interactions. Methods in Molecular Biology, vol 1421. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3591-8_7

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  • DOI: https://doi.org/10.1007/978-1-4939-3591-8_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3589-5

  • Online ISBN: 978-1-4939-3591-8

  • eBook Packages: Springer Protocols

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