Wisconsin Alumni Research Foundation

Northwestern University

Christopher A Bradfield

Yi Zhong Gu

Kristin M Dolwick

John B Hogenesch

Frank Hallock Ebetino

Andrew S Kende

Edward N Glover

Alan Poland

Lucy A Carver

Elizabeth E Dunham

Alan P Poland

The present invention provides isolated nucleic acids and proteins that are new and distinct members of the bHLH-PAS superfamily of transcription regulators. These 'MOPs' (members of PAS) are useful in a variety of research, diagnostic and therapeutic applications. Several of the MOPs of the present invention are &#x3b1;-class hypoxia-inducible factors. Several other of the MOPs of the invention are involved in circadian signal transduction.

cDNAs and proteins involved in hypoxia, circadian and orphan signal transduction pathways, and methods of use

Improved cellular assay systems for detecting polycyclic aromatic hydrocarbons, dioxins, PCBs, and other substances which are agonists of the aryl hydrocarbon receptor (AHR) are disclosed. The assays utilize one or more additional cellular proteins involved in the AHR signaling pathway, which improve the sensitivity and maximal responsiveness of the assay systems.

Sensitive bioassay for detecting agonists of the aryl hydrocarbon receptor

Murine and human Ah receptor cDNAs are provided. These molecules can be used to generate large quantities of Ah-receptor protein. The Ah receptor can be inserted into cell systems such as yeast or mammalian, expressed, and used in assays to detect agonists to the Ah receptor. The Ah receptor inserted into such systems can be either the full Ah receptor, the receptor containing deletions at its amino and carboxyl ends, or a chimeric receptor. The chimeric Ah receptor has its binding and dimerization domains replaced with an analogous region from another binding protein.

Biological assay for detecting agonists to the Ah receptor

Murine and human Ah-receptor cDNAs are provided. These molecules can be used to generate large quantities of Ah-receptor protein for use in competitive binding assays for detecting environmental pollutants. Also, the murine and human cDNAs can be used in the generation of recombinant organisms that can serve as biomonitors for environmental pollutants or in regulating gene expression for receptor agonists. Additionally, the cDNAs can be used to detect human and wildlife populations that have high susceptibility to environmental pollutants and polyacrylic aromatic hydrocarbons.

Nucleic acids encoding murine and human Ah receptors

A screening assay for recognizing the presence of dioxins (and other related toxins) in a sample is disclosed. In one aspect of the invention, Ah receptor from mice and a radioactively labelled halogenated dioxin are used in a competitive binding assay to test for the presence of toxins. The label is .sup.125 I substituted directly on the main dioxin structure. The relative binding of the toxin in the samples (in competition with labelled dioxin) for Ah receptor can be compared against standard curves. A kit is provided for running such an assay and a preferred .sup.125 I ligand is provided.

Growing community of inventors

Madison, WI, United States of America

Christopher A Bradfield