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CUTTING EDGE |
Department of Immunology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44122
In the present study we show that IL-10-mediated inhibition of
inflammatory gene expression can be mediated by an AU-rich element
(ARE) cluster present in the 3' untranslated region (3'UTR) of
sensitive genes. A series of chloramphenicol acetyl transferase (CAT)
reporter gene constructs were prepared in which different fragments
from the IL-10-sensitive KC mRNA 3'UTR were placed downstream of
the coding region of the reporter gene CAT. CAT mRNA containing the KC
3'UTR was markedly destabilized as compared with the control CAT mRNA,
and the decay rate was further increased in cells stimulated with
IL-10. The KC 3'UTR contains an ARE cluster and three isolated ARE
motifs. The ARE cluster spanning nucleotides 378–399 appeared to be
both necessary and sufficient to mediate sensitivity to IL-10 because a
116-nucleotide fragment that contains the cluster conferred
sensitivity, while mutation of the sequence between positions 378 and
399 eliminated sensitivity. The destabilizing effect of IL-10 was
relatively selective, as the stability of chimeric CAT mRNAs was not
modulated in cells treated with IFN-
or IL-4.
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