Differentiation-specific element: a cis-acting developmental switch required for the sustained transcriptional expression of the angiotensinogen gene during hormonal-induced differentiation of 3T3-L1 fibroblasts to adipocytes

RANTES is a chemoattractant cytokine (chemokine) whose gene is expressed immediately after stimulation of several cell types but upregulated late (3 to 5 days) after activation in normal T lymphocytes. Here we describe two cis-acting elements in the human RANTES promoter that act in T lymphocytes. One site interacts with NFIL6, which is activated within the first 24 h after T-cell activation. The second site binds an apparently novel complex that is upregulated later, between days 3 and 5. These data provide an explanation for the immediate-early expression of RANTES in some cell types and identify apparently novel factors contributing to late RANTES transcription in T cells. The results reveal a developmental switch occurring during normal T-cell maturation coincident with the onset of terminal differentiation and the binding of late-acting factors to sequences of the RANTES promoter.

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The deadenylation conferred by the 3' untranslated region of a developmentally controlled mRNA in Xenopus embryos is switched to polyadenylation by deletion of a short sequence element

Molecular and Cellular Biology ◽

10.1128/mcb.14.3.1893-1900.1994 ◽

1994 ◽

Vol 14 (3) ◽

pp. 1893-1900

Author(s):

P Bouvet ◽

F Omilli ◽

Y Arlot-Bonnemains ◽

V Legagneux ◽

C Roghi ◽

...

Keyword(s):

Untranslated Region ◽

Mature Oocyte ◽

Sequence Information ◽

Cellular Mechanisms ◽

Sequence Element ◽

Cis Acting ◽

Xenopus Embryos ◽

Chimeric Rnas ◽

Chimeric Transcripts ◽

Developmental Switch

The maternal Xenopus Eg mRNAs are adenylated and translated in the mature oocyte and then, after fertilization, are deadenylated and released from polysomes. Therefore, after fertilization, a change occurs in the cellular mechanisms that control mRNA adenylation. In the study reported here, we show that the 3' untranslated region of Eg2 mRNA contains a cis-acting element that is required for the deadenylation of chimeric RNAs after fertilization. This cis-acting element is contained within a single 17-nucleotide portion of the Eg2 mRNA. Disruption of this deadenylation element allows adenylation of the chimeric transcripts in the embryo. Therefore, this cis-acting element is part of the sequence information required for the developmental switch from adenylation to deadenylation of the maternal Eg2 mRNA in Xenopus embryos.

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An inducible 50-kilodalton NF kappa B-like protein and a constitutive protein both bind the acute-phase response element of the angiotensinogen gene

Molecular and Cellular Biology ◽

10.1128/mcb.10.3.1023-1032.1990 ◽

1990 ◽

Vol 10 (3) ◽

pp. 1023-1032

Author(s):

D Ron ◽

A R Brasier ◽

K A Wright ◽

J E Tate ◽

J F Habener

Keyword(s):

Molecular Mass ◽

Conditioned Medium ◽

Acute Phase ◽

Acute Phase Response ◽

Phase Response ◽

Nuclear Factor ◽

Nf Kappa B ◽

Base Pairs ◽

Cis Acting ◽

Angiotensinogen Gene

The rat angiotensinogen gene is induced in the course of the hepatic acute-phase response. We demonstrate that monocyte conditioned medium can stimulate transcription of a stably introduced reporter construct driven by 615 base pairs of the angiotensinogen 5'-flanking sequence, as well as the endogenous gene, in Reuber H35 cells. Point mutations of a cis-acting element, located 545 base pairs from the transcription start site and sharing sequence identity with known nuclear factor kappa B (NF kappa B)-binding sites, led to loss of cytokine inducibility. When cloned upstream of a minimal promoter, this cis-acting element imparted transcriptional inducibility by monocyte conditioned medium, interleukin-1, and tumor necrosis factor on a luciferase reporter gene in HepG2 cells. Two distinct proteins bound this element in vitro: a heat-stable, constitutively present, hepatic nuclear protein that gave rise to a DNase I-protected footprint covering the functionally defined element; and a binding protein of different mobility, induced by monocyte conditioned medium, which also recognized the NF kappa B-binding site of the murine kappa light-chain enhancer. UV cross-linking showed this inducible protein to have an apparent molecular mass of 50 kilodaltons, similar to that described for NF kappa B and distinct from the constitutively present protein that was shown by Southwestern (DNA-protein) blot to have a molecular mass of 32 kilodaltons. Methylation interference analysis showed that the induced species made contact points with guanine residues in the NF kappa B consensus sequence typical of NF kappa B. Induction of this binding activity did not require new protein synthesis, and 12-O-tetradecanoylphorbol-13-acetate could mimic the induction by cytokines. We thus provide direct evidence for involvement of NF kappa B or a similar factor in the hepatic acute-phase response and discuss the potential role of the presence of a constitutive nuclear factor binding the same cis element.

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An inducible 50-kilodalton NF kappa B-like protein and a constitutive protein both bind the acute-phase response element of the angiotensinogen gene.

Molecular and Cellular Biology ◽

10.1128/mcb.10.3.1023 ◽

1990 ◽

Vol 10 (3) ◽

pp. 1023-1032 ◽

Cited By ~ 64

Author(s):

D Ron ◽

A R Brasier ◽

K A Wright ◽

J E Tate ◽

J F Habener

Keyword(s):

Molecular Mass ◽

Conditioned Medium ◽

Acute Phase ◽

Acute Phase Response ◽

Phase Response ◽

Nuclear Factor ◽

Nf Kappa B ◽

Base Pairs ◽

Cis Acting ◽

Angiotensinogen Gene

The rat angiotensinogen gene is induced in the course of the hepatic acute-phase response. We demonstrate that monocyte conditioned medium can stimulate transcription of a stably introduced reporter construct driven by 615 base pairs of the angiotensinogen 5'-flanking sequence, as well as the endogenous gene, in Reuber H35 cells. Point mutations of a cis-acting element, located 545 base pairs from the transcription start site and sharing sequence identity with known nuclear factor kappa B (NF kappa B)-binding sites, led to loss of cytokine inducibility. When cloned upstream of a minimal promoter, this cis-acting element imparted transcriptional inducibility by monocyte conditioned medium, interleukin-1, and tumor necrosis factor on a luciferase reporter gene in HepG2 cells. Two distinct proteins bound this element in vitro: a heat-stable, constitutively present, hepatic nuclear protein that gave rise to a DNase I-protected footprint covering the functionally defined element; and a binding protein of different mobility, induced by monocyte conditioned medium, which also recognized the NF kappa B-binding site of the murine kappa light-chain enhancer. UV cross-linking showed this inducible protein to have an apparent molecular mass of 50 kilodaltons, similar to that described for NF kappa B and distinct from the constitutively present protein that was shown by Southwestern (DNA-protein) blot to have a molecular mass of 32 kilodaltons. Methylation interference analysis showed that the induced species made contact points with guanine residues in the NF kappa B consensus sequence typical of NF kappa B. Induction of this binding activity did not require new protein synthesis, and 12-O-tetradecanoylphorbol-13-acetate could mimic the induction by cytokines. We thus provide direct evidence for involvement of NF kappa B or a similar factor in the hepatic acute-phase response and discuss the potential role of the presence of a constitutive nuclear factor binding the same cis element.

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Multiple Cis-Acting DNA Regulatory Elements Mediate Hepatic Angiotensinogen Gene Expression

Molecular Endocrinology ◽

10.1210/mend-3-6-1022 ◽

1989 ◽

Vol 3 (6) ◽

pp. 1022-1034 ◽

Cited By ~ 32

Author(s):

Allan R. Brasier ◽

James E. Tate ◽

David Ron ◽

Joel F. Habener

Keyword(s):

Gene Expression ◽

Regulatory Elements ◽

Cis Acting ◽

Angiotensinogen Gene ◽

Dna Regulatory Elements

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The deadenylation conferred by the 3' untranslated region of a developmentally controlled mRNA in Xenopus embryos is switched to polyadenylation by deletion of a short sequence element.

Molecular and Cellular Biology ◽

10.1128/mcb.14.3.1893 ◽

1994 ◽

Vol 14 (3) ◽

pp. 1893-1900 ◽

Cited By ~ 39

Author(s):

P Bouvet ◽

F Omilli ◽

Y Arlot-Bonnemains ◽

V Legagneux ◽

C Roghi ◽

...

Keyword(s):

Untranslated Region ◽

Mature Oocyte ◽

Sequence Information ◽

Cellular Mechanisms ◽

Sequence Element ◽

Cis Acting ◽

Xenopus Embryos ◽

Chimeric Rnas ◽

Chimeric Transcripts ◽

Developmental Switch

The maternal Xenopus Eg mRNAs are adenylated and translated in the mature oocyte and then, after fertilization, are deadenylated and released from polysomes. Therefore, after fertilization, a change occurs in the cellular mechanisms that control mRNA adenylation. In the study reported here, we show that the 3' untranslated region of Eg2 mRNA contains a cis-acting element that is required for the deadenylation of chimeric RNAs after fertilization. This cis-acting element is contained within a single 17-nucleotide portion of the Eg2 mRNA. Disruption of this deadenylation element allows adenylation of the chimeric transcripts in the embryo. Therefore, this cis-acting element is part of the sequence information required for the developmental switch from adenylation to deadenylation of the maternal Eg2 mRNA in Xenopus embryos.

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Two distinct osteoblast-specific cis-acting elements control expression of a mouse osteocalcin gene.

Molecular and Cellular Biology ◽

10.1128/mcb.15.4.1858 ◽

1995 ◽

Vol 15 (4) ◽

pp. 1858-1869 ◽

Cited By ~ 441

Author(s):

P Ducy ◽

G Karsenty

Keyword(s):

Cell Line ◽

Cell Lines ◽

Osteoblastic Cell ◽

Specific Gene ◽

Osteoblastic Cell Line ◽

Specific Element ◽

Cis Acting ◽

Nuclear Extracts ◽

Osteocalcin Gene ◽

Primary Osteoblasts

Osteoblasts are cells of mesodermal origin that play a pivotal role during bone growth and mineralization. The mechanisms governing osteoblast-specific gene expression are still unknown. To understand these mechanisms, we analyzed the cis-acting elements of mouse osteocalcin gene 2 (mOG2), the best-characterized osteoblast-specific gene, by DNA transfection experiments in osteoblastic and nonosteoblastic cell lines and by DNA-binding assays. 5' deletion analysis of an mOG2 promoter-luciferase chimeric gene showed that a region located between -147 and -34 contained most if not all of the regulatory elements required for osteoblast-specific expression. Three different binding sites, called A, B, and C, for factors present in nuclear extracts of osteoblasts were identified in this short promoter by DNase I footprint assays. In gel retardation assays, the A element, located between bp -64 and -47, bound a factor present only in nuclear extracts of osteoblastic cell lines and nonmineralizing primary osteoblasts. The B element, located between bp -110 and -83, bound a ubiquitously expressed factor. The C element, located between bp -146 and -132, bound a factor present only in nuclear extracts of osteoblastic cell lines and nonmineralizing and mineralizing primary osteoblasts. When cloned upstream of a minimum osteocalcin promoter or a heterologous promoter, multimers of the A element strongly increased the activities of these promoters in osteoblastic cell lines at two different stages of differentiation but in no other cell line; we named this element osteocalcin-specific element 1 (OSE1). Multimers of the C element increased the activities of these promoters predominantly in a differentiated osteoblastic cell line; we named this element OSE2. This study demonstrates that two distinct cis-acting elements are responsible for osteoblast expression of mOG2 and provides for the first time a functional characterization of osteoblast-specific cis-acting elements. We speculate that these two elements may be important at several stages of osteoblast differentiation.

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