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Mechanisms regulating the mRNA levels of interleukin-5 and two other
coordinately expressed lymphokines in the murine T lymphoma EL4.23
H Naora and IG Young
Division of Biochemistry and Molecular Biology, John Curtin School of
Medical Research, Australian National University, Canberra.
The mechanisms that regulate the mRNA levels of interleukin-5 (IL-5) were
compared with those regulating the mRNA levels of two other coordinately
expressed lymphokines in the murine T lymphoma EL4.23. Our results indicate
that IL-5 mRNA levels are independently regulated from those of IL-2 and
granulocyte-macrophage colony-stimulating factor (GM- CSF) mRNAs. The
induction of IL-5 mRNA by phorbol 12-myristate 13- acetate (PMA)
stimulation was found to be cyclosporin A-resistant, in contrast to the
induction of IL-2 and GM-CSF mRNAs. Although the three lymphokine mRNAs
were not detected in unstimulated cells by Northern blot analysis, the
GM-CSF gene was found by nuclear run-off analysis to be constitutively
transcribed. However, the IL-2 and IL-5 genes were transcriptionally
inactive in the absence of PMA stimulation. The induction of IL-5 mRNA by
PMA stimulation primarily involved increased transcriptional activity. In
contrast, GM-CSF mRNA induction predominantly involved enhanced mRNA
stability. Both transcriptional and mRNA stabilization mechanisms appeared
to regulate IL-2 mRNA induction. The activation of IL-2 and IL-5 gene
transcription was dependent on de novo protein synthesis. Cellular
treatment with cycloheximide enhanced IL-2 gene transcription once
activation was initiated, implicating the involvement of a labile
repressor(s). Furthermore, IL-5 mRNA was more stable than IL-2 and GM-CSF
mRNAs. These latter two species were stabilized by cycloheximide,
suggesting that a labile mechanism may regulate their degradation.
Volume 83,
Issue 12,
pp. 3620-3628,
06/15/1994
Copyright © 1994 by The American Society of Hematology
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