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Regulation of AP-1 expression and activity in antigen-stimulated mast
cells: the role played by protein kinase C and the possible involvement of
Fos interacting protein
I Lewin, H Nechushtan, Q Ke and E Razin
Institute of Biochemistry, Hebrew University, Hadassah Medical School,
Jerusalem, Israel.
We have recently observed that protein kinase C (PKC) was involved in the
regulation of the accumulation of mRNAs of the AP-1 components in cultured
Abelson-transformed murine fetal-liver-derived mast cells stimulated by
exocytotic stimuli. Here we analyzed the probable regulatory effect of PKC
on the synthesis and DNA-binding activity of AP-1 complexes in immunologic
stimulated mast cells. In this study we used the interleukin-3--dependent
murine fetal-liver--derived mast cells that were not transformed by the
Abelson oncogene. Study of PKC- depleted cells showed PKC dependency of
c-fos mRNA accumulation and protein expression in IgE-Ag stimulated cells.
In contrast, the c-jun mRNA accumulation was unaffected by PKC depletion,
whereas its protein expression was dependent on this enzymatic activity.
This suggests the involvement of PKC in the regulation of translation of
c-Jun, a level of c-Jun regulation that was not previously described. The
amount of AP- 1 DNA-bound complex was also lowered in PKC-depleted cells.
Therefore, PKC plays an important regulatory role in different stages of
the signal transduction pathway because of IgE-Ag stimulation.
Surprisingly, we have observed that although the amount of total
synthesized c-Fos began to increase 15 minutes after immunologic
stimulation, the amount of c-Fos associated with Juns did not increase,
even after 45 minutes. This association was not affected by PKC. Using a
Fos-interacting protein (FIP)-cDNA probe, an expression of 2.9 kb mRNA was
detected in these cells. Furthermore, immunologic stimulation caused an
increase in the amount of a Fos-containing protein complex that bound to an
FIP-binding DNA oligonucleotide. Therefore, we propose that this protein
complex that contains most of the immunologically induced c-Fos has an
important role in IgE-Ag-stimulated signal transduction.
Volume 82,
Issue 12,
pp. 3745-3751,
12/15/1993
Copyright © 1993 by The American Society of Hematology
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