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Reconstitution and analysis of the multienzyme Escherichia coli RNA degradosome.

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.

SUBMITTER: Worrall JA

PROVIDER: S-EPMC7611026 | biostudies-literature | 2008 Oct

REPOSITORIES: biostudies-literature

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Reconstitution and analysis of the multienzyme Escherichia coli RNA degradosome.

Worrall Jonathan A R JA Górna Maria M Crump Nicholas T NT Phillips Lara G LG Tuck Alex C AC Price Amanda J AJ Bavro Vassiliy N VN Luisi Ben F BF

Journal of molecular biology 20080727 4

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 auxiliar ...[more]

PMID: 18691600

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