Deletion analysis of the Schistosoma mansoni 28-kDa glutathione S-transferase gene promoter in mammalian cells--importance of a proximal activator-protein-1 site.
The 1241-bp promoter region of the Schistosoma mansoni 28-kDa glutathione S-transferase gene (Sm28GST) was sequentially deleted and analyzed using the luciferase reporter gene system in different cell lines. The activator protein-1 (AP-1) site located at -231 seems to be responsible for the major part of the promoter activity. The 1241-bp Sm28GST promoter was not, in transient transfection experiments, activated by reagents generating reactive oxygen species, such as hydrogen peroxide (H2O2), 3-methylcholanthrene, and ter-methylhydroquinone, but was significantly stimulated by phorbol 12-myristate 13-acetate, a potent protein kinase C activator. The involvement of the -231 AP-1 site in phorbol 12-myristate 13-acetate stimulation was demonstrated. Moreover, evidence for in vitro and in vivo binding of the -231 AP-1 site to Jun/Fos dimers was obtained using mobility gel shift assays and co-transfection of embryonic F9 cells with Jun/Fos expression plasmids, respectively. The presence in S. mansoni nuclear extracts of components with affinity for the AP-1 site suggests conservation of this regulatory pathway in the parasite.
Serra
E
E
Unité INSERM 167, Institut Pasteur, Lille, France.
Zemzoumi
K
K
Dissous
C
C
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D017353:Gene Deletion; D017238:Genes, Helminth; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics*; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014162:Transfection
D017931:DNA Primers; D018509:DNA, Helminth; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D018808:Transcription Factor AP-1; D008156:Luciferases; D005982:Glutathione Transferase