What is a transactivation assay?

What is a transactivation assay?

This assay system involves a root ground tissue-specific GAL4/UAS transactivation expression system in combination with fluorescent reporter proteins. In this system, mCherry, a red fluorescent protein, can move cell to cell via diffusion, while mCherry-H2B is tightly cell autonomous.

What is transactivation of a gene?

In the context of gene regulation: transactivation is the increased rate of gene expression triggered either by biological processes or by artificial means, through the expression of an intermediate transactivator protein.

What is a transactivator of transcription?

The transactivator of transcription (TAT) protein transduction domain is an 11-amino acid positively charged peptide that has been shown to pull diverse molecules across cell membranes in vitro and in vivo.

What is transactivation protein?

HIV-1 Tat (transactivator protein) is a crucial non-structural protein of human immunodeficiency virus (HIV). The function of Tat is to bind to the viral long terminal repeat (LTR) and activate cellular transcription machinery to initiate transcription of the viral proteins.

What do coactivators do?

A coactivator is a type of transcriptional coregulator that binds to an activator (a transcription factor) to increase the rate of transcription of a gene or set of genes. The activator contains a DNA binding domain that binds either to a DNA promoter site or a specific DNA regulatory sequence called an enhancer.

How does luciferase reporter assay work?

A Luciferase Reporter Assay. When this protein activates transcription, the cell will produce luciferase enzyme. After the addition of a lysis buffer and a substrate, a luminometer quantifies the luciferase activity. If your protein activates the expression of the target gene, the amount of signal produced increases.

What is a transactivation system?

Transactivation refers to the increased rate of transcription. It can be stimulated by natural or by artificial means. By natural means, transactivation can be stimulated by endogenous transactivators, e.g. cellular or viral proteins.

What are coactivators and corepressors?

Corepressors can be enzymes that methylate or deacetylate histones causing inhibition of transcription. Coactivators are proteins that acetylate histones and phosphorylate histone H1b so that it dissociates from the receptor-binding region of DNA.

Where do coactivators bind to?

Why is luciferase used as a reporter gene?

The luciferase reporter assay is commonly used as a tool to study gene expression at the transcriptional level. It is widely used because it is convenient, relatively inexpensive, and gives quantitative measurements instantaneously.

What is the function of transactivation domain?

The transactivation domain or trans-activating domain (TAD) is a transcription factor scaffold domain which contains binding sites for other proteins such as transcription coregulators. These binding sites are frequently referred to as activation functions (AFs). TADs are named after their amino acid composition.

What are the cell-based transactivation assays?

Cell based transactivation assays employing the full-length receptors and native promoters identify both direct and indirect activators of either or both human PXR and CAR. Transactivation Assays that Identify Indirect and Direct Activators of Human Pregnane X Receptor (PXR, NR1I2) and Constitutive Androstane Receptor (CAR, NR1I3)

What is a transient transactivation assay and how is it used?

A transient transactivation assay using protoplasts derived from Arabidopsis T87ÔÇÉcultured cells is a very powerful method for characterization and functional analysis of TFs. The vector systems that we generally use in our laboratory are well described in Fujita et al. (2005).

Is there a transactivation-based assay for the detection of intercellular protein movement?

GAL4 transactivation-based assay for the detection of selective intercellular protein movement Methods Mol Biol. 2015;1217:231-43.doi: 10.1007/978-1-4939-1523-1_15. Authors Dhinesh Kumar 1 , Huan Chen, Yeonggil Rim, Jae-Yean Kim Affiliation

What is transactivation in biology?

In the context of gene regulation: transactivation is the increased rate of gene expression triggered either by biological processes or by artificial means, through the expression of an intermediate transactivator protein.