Serotonin-dependent collagenase transcription in myometrial cells requires extended AP-1 site.
Primary cultures of uterine smooth muscle cells from post-partum rats express interstitial collagenase in response to serotonin and the serotonin-dependent production of interleukin-1 (IL-1) [Wilcox, B.D., Dumin, J.A. and Jeffrey, J.J. Serotonin regulation of interleukin-1 messenger RNA in rat uterine smooth muscle cells. Relationship to the production of interstitial collagenase J. Biol. Chem., 269, (1994a), 29658]. Transient transfections of these cells indicate that rat collagenase transcription is regulated via a proximal consensus AP-1 site within an extended palindrome. Mutation of either the AP-1 site or extended palindrome (EP) decreases promoter activity to approximately 30% of the wild-type. Electrophoretic mobility shift assays reveal the binding of smooth muscle cell nuclear proteins to the AP-1 EP. This binding is barely detectable after mutation of the EP and is completely eliminated by mutation of the AP-1 heptamer. Competition experiments demonstrate that binding to the AP-1 EP is specific and of higher affinity than binding to oligonucleotides containing a mutated EP. Binding to the AP-1 EP is higher when smooth muscle cells are cultured in the presence of serotonin than in its absence. Although IL-1 is required for collagenase transcription, binding to the AP-1 EP appears to be IL-1-independent. FosB, Fra-2, c-Jun, JunB and, most abundantly, JunD bind the AP-1 EP in the absence and presence of serotonin. In contrast, Fra-1 expression and binding are serotonin-dependent suggesting that the activation of Fra-1 may be a key component of collagenase transcriptional activation.
Wilcox
C B
CB
Department of Biochemistry and Molecular Biology, A-10, Albany Medical College, 47 New Scotland Avenue, Albany, New York 12208, USA.
Weisberg
E
E
Dumin
J A
JA
Wilcox
B D
BD
Jeffrey
J J
JJ
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D017364:Collagenases / Q000235:genetics*; D005260:Female; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009215:Myometrium / Q000166:cytology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D016760:Proto-Oncogene Proteins c-fos / Q000187:drug effects / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D012701:Serotonin / Q000494:pharmacology*; D018808:Transcription Factor AP-1; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D000099169:Fos-Related Antigen 1
D017364:Collagenases; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D012701:Serotonin; D018808:Transcription Factor AP-1; D000099169:Fos-Related Antigen 1