ETS-1 transcription factor binds cooperatively to the palindromic head to head ETS-binding sites of the stromelysin-1 promoter by counteracting autoinhibition

David Baillat, Agnès Bègue, Dominique Stéhelin, Marc Aumercier

Research output: Contribution to journalArticle

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Abstract

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 photocross-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.

Original languageEnglish (US)
Pages (from-to)29386-29398
Number of pages13
JournalJournal of Biological Chemistry
Volume277
Issue number33
DOIs
StatePublished - Aug 16 2002
Externally publishedYes

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Matrix Metalloproteinase 3
Transcription Factors
Binding Sites
Transcriptional Activation
Tissue homeostasis
Genes
Electrophoretic mobility
Surface Plasmon Resonance
Gene Expression Regulation
Surface plasmon resonance
Metalloproteases
Electrophoretic Mobility Shift Assay
Pathologic Processes
Matrix Metalloproteinases
Gene expression
Transfection
Exons
Assays
Protein Isoforms
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

ETS-1 transcription factor binds cooperatively to the palindromic head to head ETS-binding sites of the stromelysin-1 promoter by counteracting autoinhibition. / Baillat, David; Bègue, Agnès; Stéhelin, Dominique; Aumercier, Marc.

In: Journal of Biological Chemistry, Vol. 277, No. 33, 16.08.2002, p. 29386-29398.

Research output: Contribution to journalArticle

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