Reconstitution and analysis of the multienzyme Escherichia coli RNA degradosome

JOURNAL OF MOLECULAR BIOLOGY

卷:382

Article

Worrall, Jonathan A. R.¹  Gorna, Maria¹  Crump, Nicholas T.¹  Phillips, Lara G.¹  Tuck, Alex C.¹  Price, Amanda J.¹  Bavro, Vassiliy N.¹  Luisi, Ben F.¹ 

[1] Univ Cambridge, Dept Biochem, Cambridge CB2 1GA, England

关键词: RNA degradosome;    RNase E;    DEAD box helicase;    polynucleotide;    phosphorylase;    Hfq;   

DOI  :  10.1016/j.jmb.2008.07.059

来源: Elsevier

【 摘 要 】

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. (C) 2008 Elsevier Ltd. All rights reserved.

【 授权许可】

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