Reconstitution and Analysis of the Multienzyme Escherichia coli RNA Degradosome

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 assemb...

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Veröffentlicht in:Journal of molecular biology 2008-10, Vol.382 (4), p.870-883
Hauptverfasser: Worrall, Jonathan A.R., Górna, Maria, Crump, Nicholas T., Phillips, Lara G., Tuck, Alex C., Price, Amanda J., Bavro, Vassiliy N., Luisi, Ben F.
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Sprache:eng
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Zusammenfassung: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.
ISSN:0022-2836
1089-8638
DOI:10.1016/j.jmb.2008.07.059