Transcriptional regulation of the human cystathionine beta-synthase-1b basal promoter: synergistic transactivation by transcription factors NF-Y and Sp1/Sp3

Citation
Yb. Ge et al., Transcriptional regulation of the human cystathionine beta-synthase-1b basal promoter: synergistic transactivation by transcription factors NF-Y and Sp1/Sp3, BIOCHEM J, 357, 2001, pp. 97-105
Citations number
38
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL JOURNAL
ISSN journal
02646021 → ACNP
Volume
357
Year of publication
2001
Part
1
Pages
97 - 105
Database
ISI
SICI code
0264-6021(20010701)357:<97:TROTHC>2.0.ZU;2-F
Abstract
Cystathionine beta -synthase (CBS) catalyses the condensation of serine and homocysteine to form cystathionine, an intermediate step in the synthesis of cysteine. Human CBS encodes five distinct 5 ' non-coding exons, the most frequent termed CBS -1a and CBS -1b, each transcribed from its own unique GC-rich TATA-less promoter. The minimal transcriptional region (- 3792 to - 3667) of the CBS -1b promoter was defined by 5 ' and 3 ' -deletions, and t ransient transfections of reporter gene constructs in HepG2 cells, characte rized by CBS transcription exclusively from the -Ib promoter, Included in t his 125 bp region are 3 GC-boxes (termed GC-a, GC-b and GC-c), an inverted CAAT-box and an E-box. By gel-shift and supershift assays, binding of speci ficity protein (Sp)l and Sp3 to the GC box elements, upstream stimulatory f actor 1 (USF-1) to the E-box, and both nuclear factor (NF)-Y and an NF-1-li ke factor to the CAAT box could be demonstrated. By transient transfections and reporter gene assays in HepG2 and Drosophiln SL2 cells, a functional i nterplay was indicated between NF-Y binding to the CAAT-box, or between USF -I binding to the E-box, and Sp1/Sp3 binding to the GC-box elements. In SL2 cells, NF-Y and Sp1/Sp3 were synergistic, Furthermore, both Spl and the lo ng Sp3 isoform transactivated the CBS -1b minimal promoter: however, the sh ort Sp3 isoforms were potent repressors. These results may explain the cell - or tissue-specific regulation of CBS transcription, and clarify the bases for alterations in CBS gene expression in human disease and Downs syndrome .