Characterization of Estrogenic and Androgenic Activities for Bisphenol A-like Chemicals (BPs): In Vitro Estrogen and Androgen Receptors Transcriptional Activation, Gene Regulation, and Binding Profiles

RBP-Jkappa is a sequence-specific DNA binding protein which plays a central role in signalling downstream of the Notch receptor by physically interacting with its intracellular region. Although at least four Notch genes exist in mammals, it is unknown whether each Notch requires a specific downstream signalling molecule. Here we report isolation and characterization of a mouse RBP-Jkappa-related gene named RBP-L that is expressed almost exclusively in lung, in contrast to the ubiquitous expression of RBP-Jkappa. For simplicity, we propose to call RBP-Jkappa RBP-J. The RBP-L protein bound to a DNA sequence almost identical to that of RBP-J. Surprisingly, RBP-L did not interact with any of the known four mouse Notch proteins. Although we found that RBP-L and EBNA-2 cooperated in transcriptional activation, they did not show significantly strong protein-protein interaction that can be detected by several in vivo and in vitro assays. This is again in contrast to physical association of RBP-J with EBNA-2. Several models to explain functional interaction between RBP-L and EBNA-2 are discussed.

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Characterization of the repressed 5S DNA minichromosomes assembled in vitro with a high-speed supernatant of Xenopus laevis oocytes

Molecular and Cellular Biology ◽

10.1128/mcb.8.10.4257-4269.1988 ◽

1988 ◽

Vol 8 (10) ◽

pp. 4257-4269

Author(s):

A Shimamura ◽

D Tremethick ◽

A Worcel

Keyword(s):

Xenopus Laevis ◽

Transcriptional Activation ◽

High Speed ◽

Histone H1 ◽

Xenopus Laevis Oocytes ◽

Xenopus Laevis Oocyte ◽

5S Rna ◽

Rna Transcription

We describe an in vitro system, based on the Xenopus laevis oocyte supernatant of Glikin et al. (G. Glikin, I. Ruberti, and A. Worcel, Cell 37:33-41, 1984), that packages DNA into minichromosomes with regularly spaced nucleosomes containing histones H3, H4, H2A, and H2B but no histone H1. The same supernatant also assembles the 5S RNA transcription complex; however, under the conditions that favor chromatin assembly, transcription is inhibited and a phased nucleosome forms over the 5S RNA gene. The minichromosomes that are fully loaded with nucleosomes remain refractory to transcriptional activation by 5S RNA transcription factors. Our data suggest that this repression is caused by a nucleosome covering the 5S RNA gene and that histone H1 is not required for regular nucleosome spacing or for gene repression in this system.

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Modeling the Interaction of Binary and Ternary Mixtures of Estradiol with Bisphenol A and Bisphenol AF in an In Vitro Estrogen-Mediated Transcriptional Activation Assay (T47D-KBluc)

Toxicological Sciences ◽

10.1093/toxsci/kfq156 ◽

2010 ◽

Vol 116 (2) ◽

pp. 477-487 ◽

Cited By ~ 38

Author(s):

Dieldrich S. Bermudez ◽

Leon E. Gray ◽

Vickie S. Wilson

Keyword(s):

Bisphenol A ◽

Transcriptional Activation ◽

Ternary Mixtures ◽

Activation Assay ◽

Binary And Ternary Mixtures ◽

Bisphenol Af

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Human TAFII130 Is a Coactivator for NFATp

Molecular and Cellular Biology ◽

10.1128/mcb.21.10.3503-3513.2001 ◽

2001 ◽

Vol 21 (10) ◽

pp. 3503-3513 ◽

Cited By ~ 11

Author(s):

Loree J. Kim ◽

Anita G. Seto ◽

Tuan N. Nguyen ◽

James A. Goodrich

Keyword(s):

Transcriptional Activation ◽

Biochemical Characterization ◽

Luciferase Reporter ◽

Transcription System ◽

Transcription In Vitro ◽

Necessary And Sufficient ◽

In Vitro Interaction ◽

Tfiid Complex

ABSTRACT NFATp is one member of a family of transcriptional activators that regulate the expression of cytokine genes. To study mechanisms of NFATp transcriptional activation, we established a reconstituted transcription system consisting of human components that is responsive to activation by full-length NFATp. The TATA-associated factor (TAFII) subunits of the TFIID complex were required for NFATp-mediated activation in this transcription system, since TATA-binding protein (TBP) alone was insufficient in supporting activated transcription. In vitro interaction assays revealed that human TAFII130 (hTAFII130) and itsDrosophila melanogaster homolog dTAFII110 bound specifically and reproducibly to immobilized NFATp. Sequences contained in the C-terminal domain of NFATp (amino acids 688 to 921) were necessary and sufficient for hTAFII130 binding. A partial TFIID complex assembled from recombinant hTBP, hTAFII250, and hTAFII130 supported NFATp-activated transcription, demonstrating the ability of hTAFII130 to serve as a coactivator for NFATp in vitro. Overexpression of hTAFII130 in Cos-1 cells inhibited NFATp activation of a luciferase reporter. These studies demonstrate that hTAFII130 is a coactivator for NFATp and represent the first biochemical characterization of the mechanism of transcriptional activation by the NFAT family of activators.

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In vivo binding of NF-κB to the IκBβ promoter is insufficient for transcriptional activation

Biochemical Journal ◽

10.1042/bj20060786 ◽

2006 ◽

Vol 400 (1) ◽

pp. 115-125 ◽

Cited By ~ 7

Author(s):

Bryan D. Griffin ◽

Paul N. Moynagh

Keyword(s):

Gene Regulation ◽

Transcription Factor ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcriptional Activation ◽

Nuclear Factor Κb ◽

Transcriptional Initiation ◽

Il Interleukin

Despite certain structural and biochemical similarities, differences exist in the function of the NF-κB (nuclear factor κB) inhibitory proteins IκBα (inhibitory κBα) and IκBβ. The functional disparity arises in part from variance at the level of gene regulation, and in particular from the substantial induction of IκBα, but not IκBβ, gene expression post-NF-κB activation. In the present study, we probe the differential effects of IL (interleukin)-1β on induction of IκBα and perform the first characterization of the human IκBβ promoter. A consensus NF-κB-binding site, capable of binding NF-κB both in vitro and in vivo, is found in the IκBβ gene 5′ flanking region. However, the IκBβ promoter was not substantially activated by pro-inflammatory cytokines, such as IL-1β and tumour necrosis factor α, that are known to cause strong activation of NF-κB. Furthermore, in contrast with IκBα, NF-κB activation did not increase expression of endogenous IκBβ as assessed by analysis of mRNA and protein levels. Unlike κB-responsive promoters, IκBβ promoter-bound p65 inefficiently recruits RNA polymerase II, which stalls at the promoter. We present evidence that this stalling is likely due to the absence of transcription factor IIH engagement, a prerequisite for RNA polymerase II phosphorylation and transcriptional initiation. Differences in the conformation of promoter-bound NF-κB may underlie the variation in the ability to engage the basal transcriptional apparatus at the IκBβ and κB-responsive promoters. This accounts for the differential expression of IκB family members in response to NF-κB activation and furthers our understanding of the mechanisms involved in transcription factor activity and IκBβ gene regulation.

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A Novel H2A/H4 Nucleosomal Histone Acetyltransferase in Tetrahymena thermophila

Molecular and Cellular Biology ◽

10.1128/mcb.19.3.2061 ◽

1999 ◽

Vol 19 (3) ◽

pp. 2061-2068 ◽

Cited By ~ 31

Author(s):

Reiko Ohba ◽

David J. Steger ◽

James E. Brownell ◽

Craig A. Mizzen ◽

Richard G. Cook ◽

...

Keyword(s):

Transcriptional Activation ◽

Histone Acetyltransferase ◽

Molecular Size ◽

Transcriptional Coactivator ◽

Acetylation Site ◽

Catalytically Active ◽

Single Polypeptide ◽

Transcriptional Output

ABSTRACT Recently, we reported the identification of a 55-kDa polypeptide (p55) from Tetrahymena macronuclei as a catalytic subunit of a transcription-associated histone acetyltransferase (HAT A). Extensive homology between p55 and Gcn5p, a component of the SAGA and ADA transcriptional coactivator complexes in budding yeast, suggests an immediate link between the regulation of chromatin structure and transcriptional output. Here we report the characterization of a second transcription-associated HAT activity from Tetrahymenamacronuclei. This novel activity is distinct from complexes containing p55 and putative ciliate SAGA and ADA components and shares several characteristics with NuA4 (for nucleosomal H2A/H4), a 1.8-MDa, Gcn5p-independent HAT complex recently described in yeast. A key feature of both the NuA4 and Tetrahymena activities is their acetylation site specificity for lysines 5, 8, 12, and 16 of H4 and lysines 5 and 9 of H2A in nucleosomal substrates, patterns that are distinct from those of known Gcn5p family members. Moreover, like NuA4, the Tetrahymena activity is capable of activating transcription from nucleosomal templates in vitro in an acetyl coenzyme A-dependent fashion. Unlike NuA4, however, sucrose gradient analyses of the ciliate enzyme, following sequential denaturation and renaturation, estimate the molecular size of the catalytically active subunit to be ∼80 kDa, consistent with the notion that a single polypeptide or a stable subcomplex is sufficient for this H2A/H4 nucleosomal HAT activity. Together, these data document the importance of this novel HAT activity for transcriptional activation from chromatin templates and suggest that a second catalytic HAT subunit, in addition to p55/Gcn5p, is conserved between yeast and Tetrahymena.

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Characterization of the repressed 5S DNA minichromosomes assembled in vitro with a high-speed supernatant of Xenopus laevis oocytes.

Molecular and Cellular Biology ◽

10.1128/mcb.8.10.4257 ◽

1988 ◽

Vol 8 (10) ◽

pp. 4257-4269 ◽

Cited By ~ 58

Author(s):

A Shimamura ◽

D Tremethick ◽

A Worcel

Keyword(s):

Xenopus Laevis ◽

Transcriptional Activation ◽

High Speed ◽

Histone H1 ◽

Xenopus Laevis Oocytes ◽

Xenopus Laevis Oocyte ◽

5S Rna ◽

Rna Transcription

We describe an in vitro system, based on the Xenopus laevis oocyte supernatant of Glikin et al. (G. Glikin, I. Ruberti, and A. Worcel, Cell 37:33-41, 1984), that packages DNA into minichromosomes with regularly spaced nucleosomes containing histones H3, H4, H2A, and H2B but no histone H1. The same supernatant also assembles the 5S RNA transcription complex; however, under the conditions that favor chromatin assembly, transcription is inhibited and a phased nucleosome forms over the 5S RNA gene. The minichromosomes that are fully loaded with nucleosomes remain refractory to transcriptional activation by 5S RNA transcription factors. Our data suggest that this repression is caused by a nucleosome covering the 5S RNA gene and that histone H1 is not required for regular nucleosome spacing or for gene repression in this system.

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Characterization of Estrogenic and Androgenic Activities for Bisphenol A-like Chemicals (BPs): In Vitro Estrogen and Androgen Receptors Transcriptional Activation, Gene Regulation, and Binding Profiles

Toxicological Sciences ◽

10.1093/toxsci/kfz173 ◽

2019 ◽

Vol 172 (1) ◽

pp. 23-37 ◽

Cited By ~ 11

Author(s):

Katherine E Pelch ◽

Yin Li ◽

Lalith Perera ◽

Kristina A Thayer ◽

Kenneth S Korach

Keyword(s):

Bisphenol A ◽

Transcriptional Activation ◽

Food Packaging ◽

Endocrine Disrupting Chemicals ◽

Consumer Products ◽

Production Volume ◽

Bisphenol S ◽

Androgenic Activity ◽

Data Gap

Abstract Bisphenol A (BPA) is a high production volume chemical widely used in plastics, food packaging, and many other products. It is well known that endocrine-disrupting chemicals might be harmful to human health due to interference with normal hormone actions. Recent studies report widespread usage and exposure to many BPA-like chemicals (BPs) that are structurally or functionally similar to BPA. However, the biological actions and toxicity of those BPs are still relatively unknown. To address this data gap, we used in vitro cell models to evaluate the ability of 22 BPs to induce or inhibit estrogenic and androgenic activity. BPA, Bisphenol AF (BPAF), bisphenol Z (BPZ), bisphenol C (BPC), tetramethyl bisphenol A (TMBPA), bisphenol S (BPS), bisphenol E (BPE), 4,4-bisphenol F (4,4-BPF), bisphenol AP (BPAP), bisphenol B (BPB), tetrachlorobisphenol A (TCBPA), and benzylparaben (PHBB) induced estrogen receptor (ER)α and/or ERβ-mediated activity. With the exception of BPS, TCBPA, and PHBB, these same BPs were also androgen receptor (AR) antagonists. Only 3 BPs were found to be ER antagonists. Bisphenol P (BPP) selectively inhibited ERβ-mediated activity and 4-(4-phenylmethoxyphenyl)sulfonylphenol (BPS-MPE) and 2,4-bisphenol S (2,4-BPS) selectively inhibited ERα-mediated activity. None of the BPs induced AR-mediated activity. In addition, we identify that the BPs can bind to ER or AR with varying degrees by a molecular modeling analysis. Taken together, these findings help us to understand the molecular mechanism of BPs and further consideration of their usage in consumer products.

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