A biotech company sells a "reporter assay kit" for researchers to easily find small molecules that can activate the Glucocorticoid Receptor (GR), a type of hormone receptor. In such an assay, a synthetic transcriptional activator is expressed that has a Gal4 DNA binding domain fused to a GR activation domain/ligand binding domain as shown in the figure. This activator is designed to bind to a synthetic piece of DNA, where it is upstream of the gene encoding for luciferase, an enzyme that produces bioluminescence in organisms such as fireflies. In other words, if the luciferase gene is expressed, the cell will emit light (aka "glow"). (b) GR GR Gal4 Gal4 Luciferase gene (a) What DNA sequence should be in place upstream of the luciferase gene (as indicated in dark blue in the figure) to ensure that this assay works? If one adds galactose to the cell, will you observe light emission from the cell? (c) If one adds a small molecule that activates GR (such as cortisol) to the cell, will you observe light emission from the cell? (d) GR is a type I hormone receptor. When cortisol, a known ligand for GR, is added to the cell, where in the cell do you expect to see free, unbound cortisol?

A biotech company sells a "reporter assay kit" for researchers to easily find small molecules that can activate the Glucocorticoid Receptor (GR), a type of hormone receptor. In such an assay, a synthetic transcriptional activator is expressed that has a Gal4 DNA binding domain fused to a GR activation domain/ligand binding domain as shown in the figure. This activator is designed to bind to a synthetic piece of DNA, where it is upstream of the gene encoding for luciferase, an enzyme that produces bioluminescence in organisms such as fireflies. In other words, if the luciferase gene is expressed, the cell will emit light (aka "glow"). (b) GR GR Gal4 Gal4 Luciferase gene (a) What DNA sequence should be in place upstream of the luciferase gene (as indicated in dark blue in the figure) to ensure that this assay works? If one adds galactose to the cell, will you observe light emission from the cell? (c) If one adds a small molecule that activates GR (such as cortisol) to the cell, will you observe light emission from the cell? (d) GR is a type I hormone receptor. When cortisol, a known ligand for GR, is added to the cell, where in the cell do you expect to see free, unbound cortisol?

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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