title: Transcriptional control of the multi-drug transporter ABCB1 by transcription factor Sp3 in different human tissues.
abstract: The ATP-binding cassette (ABC) transporter ABCB1, encoded by the multidrug resistance gene MDR1, is expressed on brain microvascular endothelium and several types of epithelium, but not on endothelia outside the CNS. It is an essential component of the blood-brain barrier. The aim of this study was to identify cell-specific controls on the transcription of MDR1 in human brain endothelium. Reporter assays identified a region of 500 bp around the transcription start site that was optimally active in brain endothelium. Chromatin immunoprecipitation identified Sp3 and TFIID associated with this region and EMSA (electrophoretic mobility shift assays) confirmed that Sp3 binds preferentially to an Sp-target site (GC-box) on the MDR1 promoter in brain endothelium. This result contrasts with findings in other cell types and with the colon carcinoma line Caco-2, in which Sp1 preferentially associates with the MDR1 promoter. Differences in MDR1 transcriptional control between brain endothelium and Caco-2 could not be explained by the relative abundance of Sp1:Sp3 nor by the ratio of Sp3 variants, because activating variants of Sp3 were present in both cell types. However differential binding of other transcription factors was also detected in two additional upstream regions of the MDR1 promoter. Identification of cell-specific controls on the transcription of MDR1 indicates that it may be possible to modulate multi-drug resistance on tumours, while leaving the blood brain barrier intact.
authors.affiliation: Department of Life, Health and Chemical Sciences, The Open University, Milton Keynes, Bucks, United Kingdom.
authors.forename: Radka
authors.initials: R
authors.lastname: Gromnicova
authors.forename: Ignacio
authors.initials: I
authors.lastname: Romero
authors.forename: David
authors.initials: D
authors.lastname: Male
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism*; D001812:Blood-Brain Barrier; D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D002469:Cell Separation; D042783:Endothelial Cells / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051705:Sp3 Transcription Factor / Q000378:metabolism*; D035362:Transcription Factor TFIID / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D035362:Transcription Factor TFIID; D051705:Sp3 Transcription Factor