Multiple mechanisms for the inhibition of rRNA synthesis during HL-60 leukemia cell differentiation.

Abstract

Treatment of HL-60 human promyelocytic leukemia cells with inducers of granulocytic differentiation produces a depletion of cellular rRNA, with the anthracycline antibiotics aclacinomycin A (ACM) and marcellomycin (MCM) causing a more rapid loss than dimethylsulfoxide (DMSO). This action is associated with a large reduction in RNA synthesis, which precedes any decreases in protein or DNA synthesis, and is specific for rRNA relative to total polyadenylated RNA synthesis. A 70% reduction in rRNA synthesis occurs within 20 minutes of ACM treatment and by 30 hours of DMSO exposure. Relative to the amount of 28S and 18S rRNA in the cells, there is a nearly complete depletion of the amount of 45S rRNA and other large rRNA precursors in cells treated with ACM, MCM, and the intercalating agent actinomycin D. In contrast, DMSO treatment produces a more coordinated decrease in 18S and 28S rRNA and rRNA precursors. The anthracycline antibiotics inhibited the synthesis of 5' proximal and 3' distal regions of the pre-rRNA transcript, while actinomycin D had a relatively sparing effect on the transcription of the 5' external transcribed spacer region of the gene relative to depletion of 3' transcripts. These studies demonstrate that different inducers of HL-60 differentiation have varying sites of action on rRNA synthesis and/or processing, with depletion of cellular rRNA as a common consequence.