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J. Biol. Chem., Vol. 277, Issue 33, 29386-29398, August 16, 2002
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From the CNRS Unité Mixte de Recherche 8526, Institut de
Biologie de Lille, Institut Pasteur de Lille, B.P. 447, 1 Rue
Calmette, 59021 Lille Cedex, France
Stromelysin-1 (matrix metalloproteinase-3) is a
member of the matrix metalloproteinase family. Regulation of its gene
expression is critical for tissue homeostasis. Patterns of increased
co-expression of stromelysin-1 and ETS-1 genes have been observed in
pathological processes. Stromelysin-1 promoter is transactivated by ETS
proteins through two palindromic head to head ETS-binding sites, an
unusual configuration among metalloproteinase promoters. By
using surface plasmon resonance, electrophoretic mobility shift assay,
and photo-cross-linking, we showed that full-length human ETS-1 (p51)
binds cooperatively to the ETS-binding site palindrome of the human
stromelysin-1 promoter, with facilitated binding of the second ETS-1
molecule to form an ETS-1·DNA·ETS-1 ternary complex. The
study of N-terminal deletion mutants allowed us to conclude that
cooperative binding implied autoinhibition counteraction, requiring the
245-330-residue region of the protein that is encoded by exon VII of
the gene. This region was deleted in the natural p42 isoform of ETS-1,
which was unable to bind cooperatively to the palindrome. Transient transfection experiments showed a good correlation between DNA binding
and promoter transactivation for p51. In contrast, p42 showed a poorer
transactivation, reinforcing the significance of cooperative binding
for full transactivation. It is the first time that ETS-1 was shown to
be able to counteract its own autoinhibition.
ETS-1 Transcription Factor Binds Cooperatively to the Palindromic
Head to Head ETS-binding Sites of the Stromelysin-1 Promoter by
Counteracting Autoinhibition*
,
*
This work was supported in part by Grant ARC 5601 from the
Association pour la Recherche sur le Cancer (to M. A.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of a BDI (Bouvse de Docteur Ingénieur)
fellowship from the CNRS and Région Nord-Pas de Calais.
§
To whom correspondence should be addressed. Tel.: 33-3-20-87-10-97;
Fax: 33-3-20-87-11-11; E-mail: marc.aumercier@ibl.fr.
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