Synergistic role of specificity proteins and upstream stimulatory factor 1 in transactivation of the mouse carboxylesterase 2/microsomal acylcarnitine hydrolase gene promoter

Mouse carboxylesterase 2 (mCES2), a microsomal acylcarnitine hydrolase, is thought to play some important roles in fatty acid (ester) metabolism, and it is therefore thought that the level of transcription of the mCES2 gene is under tight control. Examination of the tissue expression profiles revealed that mCES2 is expressed in the liver, kidney, small intestine, brain, thymus, lung, adipose tissue and testis. When the mCES2 promoter was cloned and characterized, it was revealed that Sp1 (specificity protein 1) and Sp3 could bind to a GC box, that USF (upstream stimulatory factor) 1 could bind to an E (enhancer) box, and that Sp1 could bind to an NFκB (nuclear factor κB) element in the mCES2 promoter. Co-transfection assays showed that all of these transcription factors contributed synergistically to transactivation of the mCES2 promoter. Taken together, our results indicate that Sp1, Sp3 and USF1 are indispensable factors for transactivation of the mCES2 gene promoter. To our knowledge, this is the first study in which transcription factors that interact with a CES2 family gene have been identified. The results of the present study have provided some clues for understanding the molecular mechanisms regulating mCES2 gene expression, and should be useful for studies aimed at elucidation of physiological functions of mCES2.

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Upstream stimulatory factor 1 transactivates the human gene promoter of the cardiac isoform of acetyl-CoA carboxylase

Archives of Biochemistry and Biophysics ◽

10.1016/j.abb.2005.10.025 ◽

2006 ◽

Vol 446 (1) ◽

pp. 91-100 ◽

Cited By ~ 8

Author(s):

Siyanda Makaula ◽

Tasneem Adam ◽

M. Faadiel Essop

Keyword(s):

Human Gene ◽

Gene Promoter ◽

Acetyl Coa Carboxylase ◽

Upstream Stimulatory Factor ◽

Acetyl Coa ◽

Upstream Stimulatory Factor 1 ◽

Stimulatory Factor

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

Biochemical Journal ◽

10.1042/bj3570097 ◽

2001 ◽

Vol 357 (1) ◽

pp. 97-105 ◽

Cited By ~ 32

Author(s):

Yubin GE ◽

Mark A. KONRAD ◽

Larry H. MATHERLY ◽

Jeffrey W. TAUB

Keyword(s):

Reporter Gene ◽

Intermediate Step ◽

Nuclear Factor ◽

Upstream Stimulatory Factor ◽

E Box ◽

Gc Box ◽

Specific Regulation ◽

Specificity Protein ◽

Upstream Stimulatory Factor 1 ◽

Stimulatory Factor

Cystathionine β-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 transient transfections of reporter gene constructs in HepG2 cells, characterized by CBS transcription exclusively from the −1b promoter. Included in this 125bp 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 specificity protein (Sp)1 and Sp3 to the GC-box elements, upstream stimulatory factor 1 (USF-1) to the E-box, and both nuclear factor (NF)-Y and an NF-1-like factor to the CAAT box could be demonstrated. By transient trans fections and reporter gene assays in HepG2 and Drosophila SL2 cells, a functional interplay was indicated between NF-Y binding to the CAAT-box, or between USF-1 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 Sp1 and the long Sp3 isoform transactivated the CBS −1b minimal promoter; however, the short 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 Down's syndrome.

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Regulation of human β2-microglobulin transactivation in hematopoietic cells

Blood ◽

10.1182/blood-2002-09-2924 ◽

2003 ◽

Vol 101 (8) ◽

pp. 3058-3064 ◽

Cited By ~ 48

Author(s):

Sam J. P. Gobin ◽

Paula Biesta ◽

Peter J. Van den Elsen

Keyword(s):

Hematopoietic Cells ◽

Myeloid Cell ◽

Cell Types ◽

Nuclear Factor Κb ◽

Upstream Stimulatory Factor ◽

Β2 Microglobulin ◽

E Box ◽

Infection And Inflammation ◽

Upstream Stimulatory Factor 1 ◽

Stimulatory Factor

Abstract β2-Microglobulin (β2m) is a chaperone of major histocompatibility complex (MHC) class I (–like) molecules that play a central role in antigen presentation, immunoglobulin transport, and iron metabolism. It is therefore of importance that β2m is adequately expressed in cells that perform these functions, such as hematopoietic cells. In this study, we investigated the transcriptional regulation of β2m in lymphoid and myeloid cell lines through a promoter containing a putative E box, Ets/interferon-stimulated response element (ISRE), and κB site. Here we show that upstream stimulatory factor 1 (USF1) and USF2 bind to the E box and regulate β2m transactivation. The nuclear factor κB (NF-κB) subunits p50 and p65 bind to the κB box and p65 transactivates β2m. Interferon regulatory factor 1 (IRF1), IRF2, IRF4, and IRF8, but not PU.1, bind to the Ets/ISRE, and IRF1 and IRF3 are strong transactivators of β2m. Together, all 3 boxes are important for the constitutive and cytokine-induced levels of β2m expression in lymphoid and myeloid cell types. As such, β2m transactivation is under the control of important transcriptional pathways, which are activated during injury, infection, and inflammation.

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