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Previously we determined that the immunoglobulin kappa 3`
enhancer (
Volume 270,
Number 17,
Issue of April 28, pp. 10304-10313, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
3` Enhancer
E3`) contains at least two functional DNA sequences
(PU.1/NF-EM5 and E2A) within its 132-base pair active core. We have
determined that the activities of these two sequences are insufficient
to account for the entire activity of the 132-base pair core. Using
site-directed linker scan mutagenesis across the core fragment we
identified several additional functional sequences. We used one of
these functional sequences to screen a gt11 cDNA expression
library resulting in the isolation of cDNA clones encoding the
transcription factors ATF-1 (activating transcription factor) and CREM
(cyclic AMP response element modulator). Because ATF-1 and CREM are
known to bind to cAMP response elements (CRE), this functional sequence
was named the
E3`-CRE. We show that dibutyryl cAMP can increase
E3` enhancer activity, and in transient expression assays ATF-1
caused a 4-5-fold increase in the activity of the core enhancer
while CREM-
expression resulted in repression of enhancer
activity. RNA analyses showed increased levels of ATF-1 mRNA during B
cell development and some changes in CREM transcript processing. By
joining various fragments of the E3` enhancer to the E3`-CRE,
we observed that the E3`-CRE can synergistically increase
transcription in association with the PU.1/NF-EM5 binding sites,
suggesting a functional interaction between the proteins that bind to
these DNA sequences. Consistent with this possibility, we found that
ATF-1 and CREM can physically interact with PU.1. The isolation of
activator and repressor proteins that bind to the E3`-CRE may
relate to previous conflicting results concerning the role of the cAMP
signal transduction pathway in gene transcription.
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