Upstream Stimulatory Factor-2 (USF2) Activity Is Required for Glucose Stimulation of L-Pyruvate Kinase Promoter Activity in Single Living Islet β-Cells

Type 1 iodothyronine deiodinase (D1), a selenoenzyme that catalyzes the bioactivation of thyroid hormone, is expressed mainly in the liver. Its expression and activity are modulated by several factors, but the precise mechanism of its transcriptional regulation remains unclear. In the present study, we have analyzed the promoter of human D1 gene (hDIO1) to identify factors that prevalently increase D1 activity in the human liver. Deletion and mutation analyses demonstrated that a forkhead box (FOX)A binding site and an E-box site within the region between nucleotides −187 and −132 are important for hDIO1 promoter activity in the liver. EMSA demonstrated that FOXA1 and FOXA2 specifically bind to the FOXA binding site and that upstream stimulatory factor (USF) specifically binds to the E-box element. Overexpression of FOXA2 decreased hDIO1 promoter activity, and short interfering RNA-mediated knockdown of FOXA2 increased the expression of hDIO1 mRNA. In contrast, overexpression of USF1/2 increased hDIO1 promoter activity. Short interfering RNA-mediated knockdown of FOXA1 decreased the expression of hDIO1 mRNA, but knockdown of both FOXA1 and FOXA2 restored it. The response of the hDIO1 promoter to USF was greatly attenuated in the absence of FOXA1. Taken together, these results indicate that a balance of FOXA1 and FOXA2 expression modulates hDIO1 expression in the liver.

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Follicle-Stimulating Hormone (FSH) Transiently Blocks FSH Receptor Transcription by Increasing Inhibitor of Deoxyribonucleic Acid Binding/Differentiation-2 and Decreasing Upstream Stimulatory Factor Expression in Rat Sertoli Cells

Endocrinology ◽

10.1210/en.2008-1261 ◽

2009 ◽

Vol 150 (8) ◽

pp. 3783-3791 ◽

Cited By ~ 15

Author(s):

Pushpa Viswanathan ◽

Michelle A. Wood ◽

William H. Walker

Keyword(s):

Protein Binding ◽

Sertoli Cells ◽

Fsh Receptor ◽

Upstream Stimulatory Factor ◽

Binding Studies ◽

Nuclear Extracts ◽

E Box ◽

Stimulatory Factor ◽

Stimulation Of

FSH acts through the FSH receptor (FSHR) to modulate cell processes that are required to support developing spermatozoa. Within the testis, only Sertoli cells possess receptors for FSH and are the major targets for this regulator of spermatogenesis. FSH stimulation of Sertoli cells for 24–48 h is known to induce Fshr mRNA expression through an E-box motif (CACGTG) located 25 bp upstream of the transcription start site. In contrast, FSH stimulation for 8 h inhibits Fshr transcription. DNA-protein binding studies performed using nuclear extracts from Sertoli cells show that protein binding to the Fshr promoter E-box was reduced 68% after 6 h of FSH stimulation but increased 191% over basal levels after 48 h of stimulation. The proteins binding to the Fshr E-box were identified as upstream stimulatory factor (USF)-1 and -2. FSH stimulation transiently decreased USF1 levels and increased the expression of the inhibitor of DNA binding/differentiation (ID)-2 repressor protein with the same kinetics as the decreased USF/E-box interactions. Overexpression of ID2 resulted in a dose-dependent decrease in USF-driven Fshr promoter activity in the MSC-1 Sertoli cell line, and ID2 inhibited USF binding to the Fshr E-box. Together, these studies suggest that stimulation of Sertoli cells with FSH transiently decreases expression of the USF1 activator and induces accumulation of the ID2 repressor, to block USF binding to the Fshr promoter and delay activation of Fshr transcription. This FSH-regulated mechanism may explain the cyclical changes in Fshr expression that occurs in Sertoli cells in vivo.

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Hepatic lipase promoter activity is reduced by the C-480T and G-216A substitutions present in the common LIPC gene variant, and is increased by Upstream Stimulatory Factor

Atherosclerosis ◽

10.1016/s0021-9150(00)00478-0 ◽

2001 ◽

Vol 154 (3) ◽

pp. 625-632 ◽

Cited By ~ 45

Author(s):

Gert-Jan Botma ◽

Adrie J.M. Verhoeven ◽

Hans Jansen

Keyword(s):

Promoter Activity ◽

Hepatic Lipase ◽

Gene Variant ◽

Upstream Stimulatory Factor ◽

The Common ◽

Stimulatory Factor

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Glucose stimulation of transmembrane chloride flux in individual pancreatic β-cells demonstrated with a microfluorometric technique

Diabetes Research and Clinical Practice ◽

10.1016/s0168-8227(00)81958-0 ◽

2000 ◽

Vol 50 ◽

pp. 148

Author(s):

E Grapengiesser ◽

M Eberhardson ◽

S Patterson

Keyword(s):

Β Cells ◽

Pancreatic Β Cells ◽

Glucose Stimulation ◽

Chloride Flux ◽

Stimulation Of

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Actin filament formation in pancreatic β-cells during glucose stimulation of insulin secretion

FEBS Letters ◽

10.1016/0014-5793(80)80966-5 ◽

1980 ◽

Vol 117 (1-2) ◽

pp. 299-302 ◽

Cited By ~ 36

Author(s):

Sara K. Swanston-Flatt ◽

L. Carlsson ◽

E. Gylfe

Keyword(s):

Insulin Secretion ◽

Actin Filament ◽

Filament Formation ◽

Β Cells ◽

Pancreatic Β Cells ◽

Glucose Stimulation ◽

Stimulation Of

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Upstream Stimulatory Factor Proteins Are Major Components of the Glucose Response Complex of the L-type Pyruvate Kinase Gene Promoter

Journal of Biological Chemistry ◽

10.1074/jbc.270.6.2640 ◽

1995 ◽

Vol 270 (6) ◽

pp. 2640-2643 ◽

Cited By ~ 81

Author(s):

Anne-Marie Lefran¸ois-Martinez ◽

Antoine Martinez ◽

Bénédicte Antoine ◽

Michel Raymondjean ◽

Axel Kahn

Keyword(s):

Pyruvate Kinase ◽

Gene Promoter ◽

Upstream Stimulatory Factor ◽

Kinase Gene ◽

Glucose Response ◽

Stimulatory Factor ◽

Response Complex

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Transcription factor USF2 is developmentally regulated in fetal lung and acts together with USF1 to induce SP-A gene expression

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00219.2002 ◽

2003 ◽

Vol 284 (6) ◽

pp. L1027-L1036 ◽

Cited By ~ 12

Author(s):

Erwei Gao ◽

Ying Wang ◽

Joseph L. Alcorn ◽

Carole R. Mendelson

Keyword(s):

Promoter Activity ◽

Fetal Lung ◽

Type Ii Cells ◽

Type Ii ◽

Upstream Stimulatory Factor ◽

Developmentally Regulated ◽

Rabbit Lung ◽

E Box ◽

Flanking Region ◽

Stimulatory Factor

Expression of the pulmonary surfactant protein A ( SP-A) gene is lung specific, developmentally regulated, and enhanced by hormones and factors that increase cAMP. We previously identified two E-box-like enhancers termed distal binding element (DBE) and proximal binding element (PBE) in the 5′-flanking region of the rabbit (r) SP-A gene that are essential for cAMP induction of rSP-A promoter activity (Gao E, Alcorn JL, and Mendelson CR. J Biol Chem 268: 19697–19709, 1993). We also found that DBE and PBE serve as binding sites for the basic helix-loop-helix-leucine zipper transcription factor, upstream stimulatory factor-1 (USF1) (Gao E, Wang Y, Alcorn JL, and Mendelson CR. J Biol Chem 272: 23398–23406, 1997). In the present study, PBE was used to screen a rabbit fetal lung cDNA expression library; a cDNA insert encoding the structurally related rabbit upstream stimulatory factor-2 (rUSF2) was isolated. The levels of rUSF2 mRNA reach peak levels in fetal rabbit lung at 28 days of gestation, in concert with the time of maximal induction of SP-A gene transcription. In yeast two-hybrid analysis, rUSF2 was found to preferentially form heterodimers, compared with homodimers, with rUSF1. Binding complexes of nuclear proteins isolated from fetal rabbit lung type II cells with the DBE and PBE were supershifted by anti-rUSF2 antibodies. Binding activity was enriched in nuclear proteins from type II cells compared with fibroblasts. Overexpression of rUSF2 in transfected lung A549 cells increased rSP-A promoter activity and acted synergistically with rUSF1. We suggest that heterodimers of USF2 and USF1 bound to two E-box elements in the SP-A gene 5′-flanking region serve a key role in developmental and hormonal regulation of SP-A gene expression in pulmonary type II cells.

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The mouse thymidylate synthase promoter: essential elements are in close proximity to the transcriptional initiation sites.

Molecular and Cellular Biology ◽

10.1128/mcb.9.9.4079 ◽

1989 ◽

Vol 9 (9) ◽

pp. 4079-4082 ◽

Cited By ~ 14

Author(s):

T Deng ◽

Y Li ◽

K Jolliff ◽

L F Johnson

Keyword(s):

Thymidylate Synthase ◽

Promoter Region ◽

Promoter Activity ◽

Initiation Site ◽

Essential Elements ◽

Site Directed Mutagenesis ◽

Upstream Stimulatory Factor ◽

Transcriptional Initiation ◽

Close Proximity ◽

Stimulatory Factor

The promoter region of the mouse thymidylate synthase gene was analyzed by deletion and site-directed mutagenesis. Elimination of an upstream Sp1 element reduced expression threefold, whereas elimination of an adenovirus upstream stimulatory factor element had little effect. All of the upstream elements that are essential for promoter activity are located within 22 nucleotides of the first transcriptional initiation site.

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An upstream stimulatory factor (USF) binding motif is critical for rat preprotachykinin-A promoter activity in PC12 cells

Biochemical Journal ◽

10.1042/bj3100401 ◽

1995 ◽

Vol 310 (2) ◽

pp. 401-406 ◽

Cited By ~ 18

Author(s):

J M Paterson ◽

C F Morrison ◽

S C Mendelson ◽

J McAllister ◽

J P Quinn

Keyword(s):

Promoter Activity ◽

Cell Model ◽

Nuclear Extract ◽

Regulatory Proteins ◽

Binding Motif ◽

Upstream Stimulatory Factor ◽

Promoter Fragment ◽

Helix Loop Helix ◽

E Box ◽

Stimulatory Factor

We demonstrate the presence of a functional E box motif in the proximal rat preprotachykinin-A (rPPT) promoter. This element (spanning nucleotides -67 to -47) exhibits the sequence 5′-CACGTG-3′ which is recognized and bound by the basic helix-loop-helix family of regulatory proteins. We also show that at least one of the factors bound to this rPPT promoter element in both HeLa and PC12 nuclear extract is the ubiquitously expressed transcription factor, the upstream stimulatory factor (USF). Mutation of this element by insertion of a 10 bp linker into the E box motif, in an rPPT promoter fragment spanning -865 to +92, destroys the ability of this promoter fragment to support reporter gene expression in a PC12 cell model of rPPT promoter activity. The data indicate that this rPPT E box element is likely to function as an important cis-regulatory domain in the rPPT promoter.

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