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EDTA removes RNase I bound Ca2+ leading to the inhibition of its dsRNase activity. (A) 0.1 μM dsRNA and various concentrations of RNase I purified in the presence (top panel) or absence (bottom panel) of DTT and EDTA (–DTT/–EDTA) were incubated without or with 0.5 mM EDTA in the reaction, as indicated below the gels. Reactions in lanes 10–18 contained 4 mM CaCl2. The quantification of the uncleaved RNA substrate is shown in (B). NE, no enzyme control.
