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J. Biol. Chem., Vol. 278, Issue 20, 17688-17700, May 16, 2003
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From the Following induction and activation of the early
growth response (Egr)-1 transcription factor in human chondrocytes,
interleukin-1
Egr-1 Mediates Transcriptional Repression of COL2A1
Promoter Activity by Interleukin-1
*
,,,,,¶
Rheumatology Division, Beth Israel Deaconess
Medical Center and New England Baptist Bone & Joint Institute, Harvard
Institutes of Medicine, Boston, Massachusetts 02115 and the
§ Department of Orthopaedic Surgery, Washington University
School of Medicine, St. Louis, Missouri 63110
(IL-1) suppresses the expression of the type II
collagen gene (COL2A1), associated with induction of Egr-1
binding activity in nuclear extracts. The COL2A1 proximal
promoter contains overlapping binding sites for Egr-1 and Sp1 family
members at 
119/112 bp and 81/74 bp. Mutations that block
binding of Sp1 and Sp3 to either site markedly reduce constitutive
expression of the core promoter. IL-1-induced Egr-1 binds strongly
to the 119/112 bp site, and mutations that block Egr-1 binding
prevent inhibition by IL-1. Cotransfection with pCMV-Egr1
potentiates the inhibition of COL2A1 promoter activity by
IL-1, whereas overexpression of dominant-negative Egr-1 mutant,
Wilm's tumor-1 (WT1)/Egr1, Sp1, or Sp3 reverses the inhibition by
IL-1. Cotransfection of pGL2-COL2/Gal4, in which we substituted the
critical residue for Egr-1 binding with a Gal4 binding domain and a
pCMV-Gal4-Egr1 chimera permits an inhibitory response to IL-1
that is reversed by overexpression of Gal4-CBP. Our results indicate
that IL-1-induced activation of Egr-1 binding is required for
inhibition of COL2A1 proximal promoter activity and
suggest that Egr-1 acts as a repressor of a constitutively expressed
collagen gene by preventing interactions between Sp1 and the general
transcriptional machinery.
*
This work was supported in part by National Institutes of
Health Grants AR45378 and AG22021 (to M. B. G.) and CA68544 (to P. E. A.) and a Biomedical Science Grant from the Arthritis
Foundation (to M. B. G.).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.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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