Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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DEAD-box RNA Helicases / chemistry
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DEAD-box RNA Helicases / genetics
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Endoribonucleases* / chemistry
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Endoribonucleases* / genetics
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Endoribonucleases* / metabolism
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Escherichia coli Proteins / chemistry*
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism*
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Genetic Vectors / genetics
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Genetic Vectors / metabolism
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Host Factor 1 Protein / chemistry
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Host Factor 1 Protein / genetics
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Host Factor 1 Protein / metabolism
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Multienzyme Complexes* / chemistry
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Multienzyme Complexes* / metabolism
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Nucleic Acid Conformation
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Phosphopyruvate Hydratase / chemistry
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Phosphopyruvate Hydratase / genetics
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Polyribonucleotide Nucleotidyltransferase* / chemistry
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Polyribonucleotide Nucleotidyltransferase* / genetics
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Polyribonucleotide Nucleotidyltransferase* / metabolism
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RNA Helicases* / chemistry
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RNA Helicases* / metabolism
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RNA Precursors / genetics
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RNA Precursors / metabolism
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RNA, Bacterial / metabolism*
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
Substances
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Escherichia coli Proteins
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Hfq protein, E coli
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Host Factor 1 Protein
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Multienzyme Complexes
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RNA Precursors
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RNA, Bacterial
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Recombinant Proteins
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degradosome
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Polyribonucleotide Nucleotidyltransferase
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Endoribonucleases
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ribonuclease E
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RhlB protein, E coli
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DEAD-box RNA Helicases
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RNA Helicases
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Phosphopyruvate Hydratase