Ns. Chatterjee et al., Molecular characterization of the 5 ' regulatory region of rat sodium-dependent multivitamin transporter gene, AM J P-CELL, 280(3), 2001, pp. C548-C555
Previous studies have demonstrated the involvement of a specialized, Na+-de
pendent carrier-mediated system for biotin uptake in mammalian intestine. T
he molecular identity of the carrier protein, the Na+-dependent multivitami
n transporter (SMVT), has recently been identified. Upon characterization o
f transcript expression in the rat intestine, four distinct transcript vari
ants (I-IV) due to heterogeneity at the 5'-untranslated region were found (
Chatterjee NS, Kumar CK, Ortiz A, Rubin SA, and Said HM. Am J Physiol Cell
Physiol 277: C605-C613, 1999). This finding raised the possibility that mul
tiple promoters may be involved in driving the transcription of the SMVT ge
ne. To test this possibility, we cloned the 5' regulatory region of the SMV
T gene by genome walking. A 6.5-kb genomic DNA fragment was identified and
sequenced. Three putative promoters (P1, P2, and P3) that were separated by
exons of the four previously identified transcript variants were, indeed,
found. P1 was found to contain multiple putative regulatory regions like GA
TA-1, AP-1, AP-2, and C/EBP, including several repeats of purine-rich regio
ns and two TATA-like elements. P2 and P3 were GC rich and also revealed the
presence of many putative regulatory elements including several SP-1 conse
nsus sequences. The functional identity of each promoter and the minimal re
gions required for its function were established by the luciferase assay fo
llowing transfection of rat-derived cultured intestinal epithelial IEC-6 ce
lls. The highest functional activity of the cloned promoters was found to b
e in the order of P1 > P2 > P3. These findings represent the first characte
rization of the 5' regulatory region of any mammalian SMVT gene and should
assist in the understanding of transcriptional regulation of this important
gene.