title: Constitutive expression of an antioxidant enzyme, glutathione S-transferase P1, during differentiation of human intestinal Caco-2 cells.
abstract: In this study, we investigated the molecular mechanism of the constitutive expression of an antioxidant enzyme, glutathione S-transferase (GST), during differentiation of human intestinal epithelial Caco-2 cells. We observed that the class π GST isozyme (GST P1) expression correlated with the increased expression of caudal-related homeodomain protein 2 (CDX2), a member of the mammalian homeobox family of transcription factors. In addition, transfection of Caco-2 cells with the human CDX2 cDNA resulted in enhanced expression of the GST P1 gene and protein. Chromatin immunoprecipitation assay demonstrated that CDX2 binds to the GST P1 promoter containing the putative consensus CDX-binding element, TTTAC, located at -247 upstream from the established site for transcription initiation. Using the dsDNA pull-down assay, it was revealed that CDX2 recognized and bound to the putative consensus CDX-binding element within the human GST P1 promoter region and that the amount of the CDX2 bound to the putative consensus CDX-binding element increased during Caco-2 cell differentiation. Furthermore, we demonstrated that CDX2 formed the transcriptional complex with Sp1 and bound to the putative consensus CDX-binding element within the human GST P1 promoter region. These data suggest that CDX2 binds to the human GST P1 promoter via complex formation with Sp1 and controls the constitutive expression of GST P1 during Caco-2 cell differentiation.
authors.affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Kusano
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Horie
authors.forename: Nozomi
authors.initials: N
authors.lastname: Morishita
authors.forename: Takahiro
authors.initials: T
authors.lastname: Shibata
authors.forename: Koji
authors.initials: K
authors.lastname: Uchida
mesh_terms: D000071616:CDX2 Transcription Factor; D018938:Caco-2 Cells; D002454:Cell Differentiation; D047369:Chromatin Immunoprecipitation; D018076:DNA, Complementary / Q000096:biosynthesis; D005786:Gene Expression Regulation; D051549:Glutathione S-Transferase pi / Q000235:genetics* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000166:cytology; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D000071616:CDX2 Transcription Factor; C491270:CDX2 protein, human; D018076:DNA, Complementary; D018398:Homeodomain Proteins; D007527:Isoenzymes; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi