Nucleic Acids Research · 2000 · 67 citations · 42 references
Life SciencesMicrobiologyMolecular BiologyGene ExpressionNucleic Acid BiochemistryRna BiologyHelicase FunctionRibosome BiogenesisRna StabilityRna StructureRna-binding ProteinsRna-protein InteractionsAssayCold StressDead-box Rna HelicasesEscherichia ColiRna Duplex StabilityRna Helicase ActivityRna Strand DisplacementRna-protein Binding
We have shown previously that CrhC is a unique member of the DEAD-box family of RNA helicases whose expression occurs specifically under conditions of cold stress. Here we show that recombinant His-tagged CrhC, purified from Escherichia coli, is an ATP-independent RNA binding protein possessing RNA-dependent ATPase activity which is stimulated most efficiently by rRNA and polysome preparations. RNA strand displacement assays indicate that CrhC possesses RNA unwinding activity that is adenosine nucleotide specific. Unwinding of partially duplexed RNA proceeds in the 5'-->3' but not the 3'-->5' direction using standard assay conditions. Immunoprecipitation and far-western analysis indicate that CrhC is a component of a multisubunit complex, interacting specifically with a 37 kDa polypeptide. We propose that CrhC unwinds cold-stabilized secondary structure in the 5'-UTR of RNA during cold stress.
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A DEAD-box RNA helicase in the Escherichia coli RNA degradosome
Béatrice Py, Christopher F. Higgins, Henry Krisch et al. · Nature · 1996 · 546 citations