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Vol. 12, No. 2, pp. 233-245, January 15, 1998
1 Tsukuba Life Science Center, The Institute of Physical
and Chemical Research (RIKEN), Tsukuba 305, Japan;
2 National
Institute for Advanced Interdisciplinary Research and
3 National Institute of Bioscience and Human Technology,
Agency of Industrial Science & Technology, MITI, Tsukuba 305, Japan;
4 Institute of Applied Biochemistry, University of Tsukuba,
Tsukuba 305, Japan;
5 Institute for Molecular Biology,
University of Zurich, Zurich, Switzerland;
6 Department of
Surgery/Oncology, University of California, Los Angeles,
School of Medicine and Jonsson Comprehensive Cancer Center, Los
Angeles, California 90024 USA;
7 Department of Pathology,
Harvard Medical School, Boston, Massachusetts 02115 USA;
8 Laboratory of Gene Regulation, Imperial Cancer Research
Fund, London WC2A 3PX, UK
Transcriptional activation of the c-jun gene is a critical
event in the differentiation of F9 cells. In our previous studies we
characterized an element [differentiation response element (DRE)] in
the c-jun promoter that is both necessary and sufficient to
confer the capacity for differentiation-dependent up-regulation. This
element binds the differentiation regulatory factor (DRF) complex, of
which one component is the adenovirus E1A-associated protein p300. We
have now identified activation transcription factor-2 (ATF-2) as a
DNA-binding subunit of the DRF complex. p300 and ATF-2 interact with
each other in vivo and in vitro. The bromodomain and the
C/H2 domain of p300 mediate the binding to ATF-2, which
in turn requires a proline-rich region between amino acids 112 and 350 for its interaction with p300. The phosphorylation of the serine
residue at position 121 of ATF-2 appears to be induced by protein
kinase C
(PKC) after treatment of cells with retinoic acid (RA)
or induction with E1A. In cotransfection assays, wild-type ATF-2
enhanced the transcription of an E2/tk-luciferase
construct, in conjunction with p300-E2. However, a mutant form of
ATF-2 with a mutation at position 121 (pCMVATF-2Ser121-Ala)
did not. These results suggest that ATF-2 and p300 cooperate in the
control of transcription by forming a protein complex that is
responsive to differentiation-inducing signals, such as RA or E1A, and
moreover, that the phosphorylation of ATF-2 by PKC is probably a
signaling event in the pathway that leads to the transactivation of the
c-jun gene in F9 cells.
[Key Words:
p300; ATF-2; PKC; c-jun; differentiation of F9 cells]
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