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Updated: Jul 22, 2026

Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture
Published on: August 8, 2013
Energy dependence of RNA degradation in rabbit reticulocytes
Abstract:
RNA degradation in rabbit reticulocytes was partially energy dependent. Reticulocytes were incubated for 10 h in Krebs-Henseleit bicarbonate buffer that was gassed with 95% O2-5% CO2 and contained glucose (30 mM) or 2-deoxyglucose (20 mM) and 2,4-dinitrophenol (0.2 mM). The rate of RNA degradation was reduced 41% in the presence of the metabolic inhibitors. When the buffer was gassed with 95% N2-5% CO2 and no substrate was added, the disappearance of RNA was decreased 55%. The cellular ATP content was depleted either by addition of the metabolic inhibitors or by incubation under anoxic conditions. ATP depletion did not modify the ratio of ribosomal subunits + monomers to polysomes. Puromycin and cycloheximide, which increased or decreased, respectively, the proportion of ribosomal RNA in subunits + monomers, blocked protein synthesis but did not alter the rate of RNA degradation. These experiments indicated that energy depletion inhibited RNA degradation by a mechanism that did not depend on the inhibition of protein synthesis. No evidence was obtained to indicate that the ratio of subunits + monomers to polysomes affected RNA degradation.
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