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Vol. 13, No. 19, pp. 2594-2603, October 1, 1999
Department of Biochemistry and Molecular Biology, University of
British Columbia, Vancouver, British Columbia V6T 1Z3 Canada
The RNA degradosome is a multiprotein complex required for the
degradation of highly structured RNAs. We have developed a method for
reconstituting a minimal degradosome from purified proteins. Our
results demonstrate that a degradosome-like complex containing RNase E,
PNPase, and RhlB can form spontaneously in vitro in the absence of all
other cellular components. Moreover, ATP-dependent degradation of the
malEF REP RNA by the reconstituted, minimal degradosome is
indistinguishable from that of degradosomes isolated from whole cells.
The Rne protein serves as an essential scaffold in the reconstitution
process; however, RNase E activity is not required. Rather, Rne
coordinates the activation of RhlB dependent on a 3'
single-stranded extension on RNA substrates. A model for
degradosome-mediated degradation of structured RNA is presented with
its implications for mRNA decay in Escherichia coli.
[Key Words: RNA degradosome; 3' exonuclease; E. coli; DEAD box; RNA helicase]
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