Transcriptional regulation of the human CD97 promoter by Sp1/Sp3 in smooth muscle cells.
The EGF-TM7 receptor CD97 shows different features of expression and function in muscle cells compared to hematopoetic and tumor cells. Since the molecular function and regulation of CD97 are poorly understood, this study aimed at defining its basal transcriptional regulation in smooth muscle cells (SMCs). The computational analysis of the CD97 5'-flanking region revealed that the TATA box-lacking promoter possesses several GC-rich regions as putative Sp1/Sp3 binding sites. Transfection studies with serially deleted promoter constructs demonstrated that the minimal promoter fragment resided in the -218/+45 region containing one out of five identified GC-boxes in the leiomyosarcoma cell line SK-LMS-1 and human bronchial smooth muscle cells (HbSMCs). Mutation of the most proximal GC-site in CD97 reporter gene constructs caused a significant decrease in promoter activity. Gel shift assays and chromatin immunoprecipitation revealed that Sp1 and Sp3 bound specifically to the most proximal GC-site. Furthermore, we showed that Sp1 and Sp3 over-expression activates CD97 promoter activity in HEK293 cells. Our data characterize for the first time the activity of the human CD97 promoter which is controlled by Sp1/Sp3 transcription factors in SMCs.
Wobus
Manja
M
Department of Surgery, Research Laboratories, University of Leipzig, Leipzig, Germany. manja.wobus@uniklinikum-dresden.de
Wandel
Elke
E
Prohaska
Sonja
S
Findeiss
Sven
S
Tschöp
Katrin
K
Aust
Gabriela
G
D015703:Antigens, CD / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D020862:GC Rich Sequence; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D043562:Receptors, G-Protein-Coupled; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D051705:Sp3 Transcription Factor / Q000378:metabolism*; D015533:Transcriptional Activation
C095004:ADGRE5 protein, human; D015703:Antigens, CD; D017931:DNA Primers; D008562:Membrane Glycoproteins; D043562:Receptors, G-Protein-Coupled; C497121:SP3 protein, human; D016329:Sp1 Transcription Factor; D051705:Sp3 Transcription Factor; D004247:DNA; D008156:Luciferases