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Identification and molecular characterization of 5′‐regulatory region of the human SLC52A1 (rft‐1) promoter (896.4)
Author(s) -
Sabui Subrata,
Said Hamid
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.896.4
Subject(s) - promoter , gene , biology , microbiology and biotechnology , transcription factor , regulatory sequence , transcription (linguistics) , tata box , genetics , gene expression , philosophy , linguistics
The human SLC52A1 gene encodes riboflavin transporter‐1 (rft‐1) which is essential for riboflavin transport across cell membranes. Rft‐1 is strongly expressed in human placenta, small intestine, heart muscle and thymus. Nothing is known about the transcriptional regulation of the human SLC52A1. We addressed the issue by cloning the 5'‐regulatory region of the SLC52A1 gene, identified the minimal promoter region for basal activity and identified potential cis‐regulatory elements. Guided by Rapid amplification of the cDNA ends (5'‐RACE), evidence was obtained for existence of one transcription start site (TSS) at nt‐716 in intestinal epithelial cells (Caco‐2). A 384‐bp segment upstream of the TSS was found to exhibit significant promoter activity in different human derived intestinal epithelial cells (Caco‐2, HuTu80 and NCM460). Truncation of this segment revealed that core promoter activity was embedded in a 297‐bp fragment (‐146 to +149) that contains GC rich, lacks TATA element and harbors several putative transcription factor binding sites includes SP‐1, AP‐2 and NFkB. Mutating the latter sites resulted in a significant decrease in promoter activity suggesting involvement of these cis‐regulatory elements in the regulation of SLC52A1 transcription. This study reports on first characterization of the SLC52A1 promoter. Grant Funding Source : Supported by the VA and NIH, DK56061