Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor–reporter gene assays

1996 ◽  
Vol 74 (2) ◽  
pp. 216-222 ◽  
Author(s):  
Patrick Balaguer ◽  
Annick Joyeux ◽  
Michael S Denison ◽  
Renaud Vincent ◽  
Bradley E Gillesby ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Complex Mixtures ◽  
Recombinant Receptor ◽  
Reporter Gene Assays

Assessing the estrogenic and dio×in-like activities of chemicals and comple× mi×tures using in vitro recombinant receptor–reporter gene assays

1996 ◽  
Vol 74 (2) ◽  
pp. 216-222 ◽  
Author(s):  
Patrick Balaguer ◽  
Annick Joyeu× ◽  
Michael S. Denison ◽  
Renaud Vincent ◽  
Bradley E. Gillesby ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Recombinant Receptor ◽  
Reporter Gene Assays

Screening for androgen receptor activities in 253 industrial chemicals by in vitro reporter gene assays using AR-EcoScreenTM cells

2005 ◽  
Vol 19 (6) ◽  
pp. 831-842 ◽  
Author(s):  
Naohiro Araki ◽  
Ken Ohno ◽  
Makoto Nakai ◽  
Masahiro Takeyoshi ◽  
Mitsuru Iida
Keyword(s):  
Androgen Receptor ◽  
Reporter Gene ◽  
Industrial Chemicals ◽  
Reporter Gene Assays

The MDS-Associated Protein EVI1 Impairs Erythroid and Megakaryocytic Differentiation by Direct Interaction with GATA-1.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3007-3007
Author(s):  
Leopoldo Laricchia-Robbio ◽  
Donglan Li ◽  
Raffaella Fazzina ◽  
Soumen Chakraborty ◽  
Maher Abdul Hay ◽  
...  
Keyword(s):  
Dna Binding ◽  
Reporter Gene ◽  
Chromosomal Abnormalities ◽  
Point Mutations ◽  
Binding Motif ◽  
Murine Bone Marrow ◽  
C Terminus ◽  
N Terminus ◽  
Reporter Gene Assays

Abstract EVI1 is an aggressive nuclear oncoprotein deregulated by recurring chromosomal abnormalities in acute myeloid leukemia and myelodysplastic syndrome. This protein has two Zn finger domains containing 7 motifs at the N-terminus and 3 motifs at the C-terminus. The expression of this gene is a very poor prognostic marker and is associated with diseases characterized by erythroid and megakaryocytic defects. We have recently shown that the forced expression of EVI1 in murine bone marrow results in a fatal disease with features characteristic of MDS, including fatal dyserythropoiesis, dysmegakaryopoiesis, and anemia. These lineages are regulated by the transcription factor GATA-1, a DNA-binding protein that in addition to erythrocytes and megakaryocytes exerts a strict control also on the differentiation of mast cells and eosinophils, on the basis of its expression and association with specific partners. In the present study, we used biochemical assays and in vitro culture to show that GATA-1 and the N-terminus of EVI1 are involved in the formation of a protein complex that is unable to regulate efficiently GATA-1-dependent promoters in reporter gene assays. EMSA studies with a GATA-1-specific probe indicate that EVI1 does not recognize and bind to the DNA probe but disrupts the DNA-binding of GATA-1. By deletion analysis and point mutations, we mapped the interaction between the proteins to two motifs in the proximal Zn finger domain of EVI1 and to the C-terminus Zn finger of GATA-1. Cys to Ala mutations in the two EVI1 motifs abrogate the interaction and restore the response of a promoter in reporter gene assays. We propose that the association between EVI1 and the DNA-binding motif of GATA-1 impairs efficient promoter binding by GATA-1 and the regulation of erythroid and megakaryocytic lineage. There studies suggest that the interaction surface between the two proteins could be an attractive target for the development of competing small molecules as a treatment in EVI1-associated leukemia.

scholarly journals Evaluation of Firefly and Renilla Luciferase Inhibition in Reporter-Gene Assays: A Case of Isoflavonoids

2021 ◽  
Vol 22 (13) ◽  
pp. 6927
Author(s):  
Maša Kenda ◽  
Jan Vegelj ◽  
Barbara Herlah ◽  
Andrej Perdih ◽  
Přemysl Mladěnka ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Firefly Luciferase ◽  
Qsar Model ◽  
In Vitro Assay ◽  
Biochanin A ◽  
Renilla Luciferase ◽  
Luciferase Reporter ◽  
Vitro Assay ◽  
Reporter Gene Assays

Firefly luciferase is susceptible to inhibition and stabilization by compounds under investigation for biological activity and toxicity. This can lead to false-positive results in in vitro cell-based assays. However, firefly luciferase remains one of the most commonly used reporter genes. Here, we evaluated isoflavonoids for inhibition of firefly luciferase. These natural compounds are often studied using luciferase reporter-gene assays. We used a quantitative structure–activity relationship (QSAR) model to compare the results of in silico predictions with a newly developed in vitro assay that enables concomitant detection of inhibition of firefly and Renilla luciferases. The QSAR model predicted a moderate to high likelihood of firefly luciferase inhibition for all of the 11 isoflavonoids investigated, and the in vitro assays confirmed this for seven of them: daidzein, genistein, glycitein, prunetin, biochanin A, calycosin, and formononetin. In contrast, none of the 11 isoflavonoids inhibited Renilla luciferase. Molecular docking calculations indicated that isoflavonoids interact favorably with the D-luciferin binding pocket of firefly luciferase. These data demonstrate the importance of reporter-enzyme inhibition when studying the effects of such compounds and suggest that this in vitro assay can be used to exclude false-positives due to firefly or Renilla luciferase inhibition, and to thus define the most appropriate reporter gene.

scholarly journals High-Throughput Effect-Directed Analysis Using Downscaled in Vitro Reporter Gene Assays To Identify Endocrine Disruptors in Surface Water

2018 ◽  
Vol 52 (7) ◽  
pp. 4367-4377 ◽  
Author(s):  
Nick Zwart ◽  
Shan Li Nio ◽  
Corine J. Houtman ◽  
Jacob de Boer ◽  
Jeroen Kool ◽  
...  
Keyword(s):  
Surface Water ◽  
Endocrine Disruptors ◽  
Reporter Gene ◽  
High Throughput ◽  
Effect Directed Analysis ◽  
Reporter Gene Assays

Comparison of in vitro hormone activities of selected phthalates using reporter gene assays

2009 ◽  
Vol 191 (1) ◽  
pp. 9-14 ◽  
Author(s):  
Ouxi Shen ◽  
Guizhen Du ◽  
Hong Sun ◽  
Wei Wu ◽  
Yi Jiang ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assays
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