TITLE

Impacts of different promoters on the mammalian one-hybrid assay for detecting nuclear receptor agonists

AUTHOR(S)
Zhi-Hui Zheng; Xin-Hua Lu; Hua Zhang; Guo-Ping Lv; Jian-Gong He; Bao-Hua Zhao; Shu-Yi Si
PUB. DATE
March 2010
SOURCE
Analytical & Bioanalytical Chemistry;Mar2010, Vol. 396 Issue 5, p1721
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Nuclear receptors are a superfamily of ligand-activated transcription factors that play key roles in many biological processes, and have become one class of the most important targets in drug discovery. Mammalian one-hybrid system has been used to develop a cell-based functional transactivation high-throughput screening (HTS) assay for detecting nuclear receptors ligands. In the present study, we proved that different promoters used in the reporter vector had significant different impacts on the performance of HTS assays. The assay using the SV40 promoter in the reporter vector showed the characteristics of much higher signal/noise ratios, acceptable Z′ factors (>0.6), low coefficient variation (<12.5%) and higher hits rate, which could be more robust, reproducible, and sensitive. In contrast, utilizing a TATA box promoter in the assay resulted in higher variance and low sensitivity. In addition, it was found that the assay using SV40 had longer signal decay time and was easier to be miniaturized in 384-well format. It has been confirmed that the choice of a promoter is a critical factor in developing a reporter gene HTS assay. However, the SV40 promoter used in the present study has been shown to be more adaptable than the minimal promoter TATA box in the Mammalian one-hybrid HTS assays for detecting nuclear receptor agonists. [Figure not available: see fulltext.]
ACCESSION #
48191139

 

Related Articles

  • Liver X Receptor: Crosstalk Node for the Signaling of Lipid Metabolism, Carbohydrate Metabolism, and Innate Immunity. Fessler, Michael B. // Current Signal Transduction Therapy;May2008, Vol. 3 Issue 2, p75 

    Liver X Receptor-a (LXRa, also known as NR1H3) and LXR� (NR1H2) are members of the nuclear receptor superfamily of ligand-activated transcription factors, a superfamily which includes the more widely known glucocorticoid receptor, estrogen receptor, thyroid receptor, and peroxisome...

  • Use of Mouse Models to Evaluate Roles of Nuclear Receptors and their Ligands in the Pathogenesis and Treatment of Atherosclerosis. Li, A. C.; Glass, C. K. // Current Drug Targets;Dec2007, Vol. 8 Issue 12, p1273 

    Nuclear receptors form a large family of ligand-dependent transcription factors that regulate diverse aspects of development and homeostasis. Several of these receptors have been demonstrated to play important roles in controlling biological processes that influence the development and clinical...

  • Non-Steroidal Steroid Receptor Modulators. Buijsman, Rogier C.; Hermkens, Pedro H. H.; van Rijn, Rachel D.; Stock, H. T.; Teerhuis, N. Miranda // Current Medicinal Chemistry;2005, Vol. 12 Issue 9, p1017 

    The last ten years much attention has been focused on the finding of non-steroidal ligands for steroidal nuclear receptors for reasons such as diminishing cross-reactivity to eliminate side effect profiles, changing physicochemical properties which might cause different tissue distribution...

  • Selection, Application, and Validation of a Set of Molecular Descriptors for Nuclear Receptor Ligands. Stewart, Eugene L.; Brown, Peter J.; Bentley, James A.; Wilison, Timothy M. // Combinatorial Chemistry & High Throughput Screening;Aug2004, Vol. 7 Issue 5, p407 

    A methodology for the selection and validation of nuclear receptor ligand chemical descriptors is described. After descriptors for a targeted chemical space were selected, a virtual screening methodology utilizing this space was formulated for the identification of potential NR ligands from our...

  • Dissecting the Relation between a Nuclear Receptor and GATA: Binding Affinity Studies of Thyroid Hormone Receptor and GATA2 on TSHβ Promoter. Figueira, Ana Carolina Migliorini; Polikarpov, Igor; Veprintsev, Dmitry; Santos, Guilherme Martins // PLoS ONE;2010, Vol. 5 Issue 9, p1 

    Background: Much is known about how genes regulated by nuclear receptors (NRs) are switched on in the presence of a ligand. However, the molecular mechanism for gene down-regulation by liganded NRs remains a conundrum. The interaction between two zinc-finger transcription factors, Nuclear...

  • Synthesis of a Novel Series of 8-HETE Analogs and their Biological Evaluation Towards the PPAR Nuclear Receptors. Liutkus, M�lanie; Dacquet, Catherine; Audinot-Bouchez, Val�rie; Boutin, Jean; Caignard, Daniel-Henri; Ktorza, Alain; Gr�e, Ren� // Letters in Drug Design & Discovery;Nov2008, Vol. 5 Issue 8, p503 

    A novel series of dual PPARa/? agonists was designed through the modification of our previously described family of 8-HETE analogs. The combination of the structural elements of this family and of the classical PPAR ligands produced compounds with a quinoxaline core and two sides chains....

  • The Pharmacology of LXR. Michael, Laura F.; Schkeryantz, Jeffrey M.; Burris, Thomas P. // Mini Reviews in Medicinal Chemistry;Aug2005, Vol. 5 Issue 8, p729 

    Liver X receptors (LXRs) are members of the nuclear hormone receptor superfamily of ligandactivated transcription factors. Two LXRs (LXRa and LXRb) were initially characterized as orphan members of this superfamily with disparate patterns of tissue expression. These two receptors later were...

  • Basis of a High-Throughput Method for Nuclear Receptor Ligands. Kanayama, Tomohiko; Mamiya, Satoru; Nishihara, Tsutomu; Nishikawa, Jun-ichi // Journal of Biochemistry;Jun2003, Vol. 133 Issue 6, p791 

    Assessment of the risk of human exposure to man-made chemicals that bind to hormone receptors has emerged as a major public health issue. Among hormone receptors, nuclear receptors tend to be targets of xenobiotics because their endogenous ligands are small, fat-soluble molecules. Nuclear...

  • Cross-Talk between PPARs and the Partners of RXR: A Molecular Perspective. Alan Chan, Lap Shu; Wells, Richard A. // PPAR Research;2009, Special section p1 

    The PPARs are integral parts of the RXR-dependent signaling networks. Many other nuclear receptor subfamily 1 members also require RXR as their obligatory heterodimerization partner and they are often co-expressed in any given tissue. Therefore, the PPARs often complete with other RXR-dependent...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics