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The Escherichia coli RNA degradosome is a multienzyme assembly that functions in transcript turnover and maturation of structured RNA precursors. We have developed a procedure to reconstitute the RNA degradosome from recombinant components using modular coexpression vectors. The reconstituted assembly can be purified on a scale that has enabled biochemical and biophysical analyses, and we compare the properties of recombinant and cell-extracted RNA degradosomes. We present evidence that auxiliary protein components can be recruited to the 'superprotomer' core of the assembly through a dynamic equilibrium involving RNA intermediaries. We discuss the implications for the regulation of RNA degradosome function in vivo.

Original publication

DOI

10.1016/j.jmb.2008.07.059

Type

Journal article

Journal

J Mol Biol

Publication Date

17/10/2008

Volume

382

Pages

870 - 883

Keywords

DEAD-box RNA Helicases, Endoribonucleases, Escherichia coli, Escherichia coli Proteins, Genetic Vectors, Host Factor 1 Protein, Multienzyme Complexes, Nucleic Acid Conformation, Phosphopyruvate Hydratase, Polyribonucleotide Nucleotidyltransferase, RNA Helicases, RNA Precursors, RNA, Bacterial, Recombinant Proteins