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