PMID- 25394472
OWN - NLM
STAT- MEDLINE
DCOM- 20150317
LR  - 20220819
IS  - 1522-1563 (Electronic)
IS  - 0363-6143 (Print)
IS  - 0363-6143 (Linking)
VI  - 308
IP  - 2
DP  - 2015 Jan 15
TI  - Identification and characterization of the minimal 5'-regulatory region of the 
      human riboflavin transporter-3 (SLC52A3) in intestinal epithelial cells.
PG  - C189-96
LID - 10.1152/ajpcell.00342.2014 [doi]
AB  - The human riboflavin (RF) transporter-3 (product of the SLC52A3 gene) plays an 
      important role in intestinal RF absorption. Our aims in this study were to 
      identify the minimal 5'-regulatory region of the SLC52A3 gene and the regulatory 
      element(s) involved in its activity in intestinal epithelial cells, as well as to 
      confirm promoter activity and establish physiological relevance in vivo in 
      transgenic mice. With the use of transiently transfected human intestinal 
      epithelial HuTu 80 cells and 5'-deletion analysis, the minimal SLC52A3 promoter 
      was found to be encoded between -199 and +8 bp (using the start of the 
      transcription start site as position 1). Although several putative cis-regulatory 
      elements were predicted in this region, only the stimulating protein-1 (Sp1) 
      binding site (at position -74/-71 bp) was found to play a role in promoter 
      activity, as indicated by mutational analysis. Binding of Sp1 to the minimal 
      SLC52A3 promoter was demonstrated by means of EMSA and supershift assays and by 
      chromatin immunoprecipitation analysis. Studies with Drosophila SL2 cells (which 
      lack Sp activity) confirmed the importance of Sp1 in driving the activity of the 
      SLC52A3 minimal promoter; they further showed that Sp3 can also do the 
      activation. Finally, with the use of luciferase gene fusions, the activity of the 
      cloned SLC52A3 promoter was confirmed in vivo in transgenic mice. These studies 
      report, for the first time, on the identification and characterization of the 
      SLC52A3 promoter and also demonstrate the importance of Sp1 in regulating its 
      activity in intestinal epithelial cells.
FAU - Ghosal, Abhisek
AU  - Ghosal A
AD  - Departments of Medicine and Physiology/Biophysics, University of California, 
      Irvine, California; and Department of Veterans Affairs Medical Center, Long 
      Beach, California.
FAU - Sabui, Subrata
AU  - Sabui S
AD  - Departments of Medicine and Physiology/Biophysics, University of California, 
      Irvine, California; and Department of Veterans Affairs Medical Center, Long 
      Beach, California.
FAU - Said, Hamid M
AU  - Said HM
AD  - Departments of Medicine and Physiology/Biophysics, University of California, 
      Irvine, California; and Department of Veterans Affairs Medical Center, Long 
      Beach, California hmsaid@uci.edu.
LA  - eng
GR  - R37 DK056061/DK/NIDDK NIH HHS/United States
GR  - R01 DK058057/DK/NIDDK NIH HHS/United States
GR  - AA018071/AA/NIAAA NIH HHS/United States
GR  - R01 AA018071/AA/NIAAA NIH HHS/United States
GR  - DK58057/DK/NIDDK NIH HHS/United States
GR  - DK56061/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20141112
PL  - United States
TA  - Am J Physiol Cell Physiol
JT  - American journal of physiology. Cell physiology
JID - 100901225
RN  - 0 (Membrane Transport Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Receptors, G-Protein-Coupled)
RN  - 0 (SLC52A2 protein, human)
RN  - 0 (SLC52A3 protein, human)
RN  - 0 (Sp1 Transcription Factor)
RN  - 148710-94-5 (Sp3 Transcription Factor)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Binding Sites/genetics
MH  - Cells, Cultured
MH  - Chromatin Immunoprecipitation/methods
MH  - Drosophila/genetics/metabolism
MH  - Epithelial Cells/*metabolism
MH  - Humans
MH  - Intestinal Mucosa/*metabolism
MH  - Luciferases/genetics/metabolism
MH  - Membrane Transport Proteins/*genetics/*metabolism
MH  - Mice
MH  - Mice, Transgenic
MH  - Molecular Sequence Data
MH  - Nerve Tissue Proteins/genetics/metabolism
MH  - Promoter Regions, Genetic/genetics
MH  - Receptors, G-Protein-Coupled
MH  - Regulatory Sequences, Nucleic Acid/*genetics
MH  - Sp1 Transcription Factor/genetics/metabolism
MH  - Sp3 Transcription Factor/genetics/metabolism
PMC - PMC4297772
OTO - NOTNLM
OT  - intestinal cells
OT  - minimal promoter region
OT  - promoter activity
OT  - riboflavin
OT  - riboflavin transporter-3
OT  - stimulating protein-1
EDAT- 2014/11/15 06:00
MHDA- 2015/03/18 06:00
PMCR- 2016/01/15
CRDT- 2014/11/15 06:00
PHST- 2014/11/15 06:00 [entrez]
PHST- 2014/11/15 06:00 [pubmed]
PHST- 2015/03/18 06:00 [medline]
PHST- 2016/01/15 00:00 [pmc-release]
AID - ajpcell.00342.2014 [pii]
AID - C-00342-2014 [pii]
AID - 10.1152/ajpcell.00342.2014 [doi]
PST - ppublish
SO  - Am J Physiol Cell Physiol. 2015 Jan 15;308(2):C189-96. doi: 
      10.1152/ajpcell.00342.2014. Epub 2014 Nov 12.