Crystallization of RNA-protein complexes: From synthesis and purification of individual components to crystals

Research output: Chapter in Book/Report/Conference proceedingChapter

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Abstract

A broad range of biological processes relies on complexes between RNA and proteins. Crystallization of RNA-protein complexes can yield invaluable information on structural organizations of key elements of cellular machinery. However, crystallization of RNA-protein complexes is often challenging and requires special approaches. Here we review the purification of RNA, RNA-binding proteins, and the formation and crystallization of RNA-protein complexes, using the crystallization of the P3 RNA domain of ribonuclease MRP, a multicomponent ribonucleoprotein complex involved in the metabolism of various RNA molecules, as an example. The RNA-protein complex was formed using gel-purified RNA, produced by run-off transcription with T7 RNA polymerase in vitro, and proteins that were overexpressed in Escherichia coli and purified to be RNase-free. The complex was crystallized using a sitting drop setup; initial screening for suitable crystallization conditions was performed using a sparse matrix approach.

Original languageEnglish (US)
Title of host publicationBacterial Regulatory RNA
Subtitle of host publicationMethods and Protocols
EditorsKenneth C. Keiler
Pages123-143
Number of pages21
DOIs
StatePublished - Jul 27 2012

Publication series

NameMethods in Molecular Biology
Volume905
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Perederina, A., & Krasilnikov, A. S. (2012). Crystallization of RNA-protein complexes: From synthesis and purification of individual components to crystals. In K. C. Keiler (Ed.), Bacterial Regulatory RNA: Methods and Protocols (pp. 123-143). (Methods in Molecular Biology; Vol. 905). https://doi.org/10.1007/978-1-61779-949-5_9