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Granulocyte-macrophage colony-stimulating factor, phorbol ester, and sodium
butyrate induce the CD11c integrin gene promoter activity during myeloid
cell differentiation
MA Rubio, C Lopez-Rodriguez, A Nueda, P Aller, AL Armesilla, MA Vega and AL Corbi
Hospital de la Princesa, Centro de Investigaciones Biologicas, Consejo
Superior de Investigaciones Cientificas (CSIC), Madrid, Spain.
To analyze the activity of the CD11c promoter during myeloid
differentiation without the limitations of transient expression systems, we
have stably transfected the myeloid U937 cell line with the pCD11C361-Luc
plasmid, in which the expression of the firefly luciferase cDNA is driven
by the CD11c promoter region -361/+43, previously shown to confer myeloid
specificity to reporter genes. The stable transfectants (U937-C361)
retained the ability to differentiate in response to phorbol-ester (PMA),
sodium butyrate (SB), granulocyte- macrophage colony-stimulating factor
(GM-CSF), and other differentiating agents. U937-C361 differentiation
correlated with increased cellular luciferase levels, showing the
inducibility of the CD11c promoter during myeloid differentiation and
establishing the U937- C361 cells as a suitable system for studying the
myeloid differentiation-inducing capacity of cytokines, growth, factors,
and other biological response modifiers. Unexpectedly, the inducibility of
the CD11c gene promoter showed distinct kinetics and magnitude on the PMA-,
SB-, GM-CSF-triggered differentiation. Moreover, SB synergized with either
PMA or GM-CSF in enhancing both the CD11c promoter activity and the cell
surface expression of p150,95 on differentiating U937 cells. Furthermore,
we showed the existence of a c-Myb-binding site at - 85, the importance of
the -99/-61 region in the CD11c promoter inducibility during PMA- or
SB-triggered differentiation, and the dependency of the GM-CSF and PMA
responsiveness of the CD11c promoter on an intact AP-1-binding site located
at -60. These results, together with the lack of functional effect of
mutations disrupting the Sp1-and Myb-binding sites within the proximal
region of the CD11c promoter, indicate that the myeloid differentiation
pathways indicated by SB and phorbol esters (or GM-CSF) activate a distinct
set of transcription factors and show that the myeloid
differentiation-inducibility of the CD11c gene maps to the -99/-53 proximal
region of the promoter.
Volume 86,
Issue 10,
pp. 3715-3724,
11/15/1995
Copyright © 1995 by The American Society of Hematology
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