1. Sponsor Organization: NIH/NIEHS

2. Project Title: 01884-20 HALOGENATED AROMATIC HYDROCARBONS

3. Project Focus:

• Project Primary Focus: Human Health Effects
• Project Secondary Focus:

4. Description:

The goal of this proposal is to characterize the Ah receptor - a soluble protein present in many tissues of vertebrate species and a presumed member of the erbA superfamily of receptors, which mediates the pleiotropic effects of halogenated aromatic hydrocarbons such as 2,3,7,8-tetrachlorodibenzo-p-dioxin. 1) We propose to clone and sequence the gene for the Ah receptor from cDNA libraries by using degenerate oligonucleotide probes and rabbit polyclonal antibodies to a synthetic peptide that corresponds to the N-terminal sequence of the receptor. We plan to analyze the structure-function analysis of this protein using mutagenesis and an expression assay. 2) We propose to characterize the four allelic variants of the Ah receptor among inbred mouse strains: a) in vitro by their binding kinetics K-D, B-MAX and ease of activation and b) in vivo by genetic analysis of the differences in functional phenotypes especially Ah(b-1)/Ah(d) vs. Ah(b-2)/Ah(d) and correlation of this with quantitative nuclear extract-gel retardation experiments. 3) We propose to expand on our photoaffinity labeling and immunologic methodology; a) to screen lower vertebrate and invertebrate species for the presence of the Ah receptor, b) to prepare new monoclonal and polyclonal antibodies to various regions of the Ah receptor, c) to prepare an immunoaffinity column to aid in rapid purification, and d) to examine murine tissues for the cellular localization of the Ah receptor using immunohistochemistry.

5. References:

6. Inventory Category:

• Primary: Models
• Secondary:

7. Inventory Subcategory:

• Primary: Basic Research
• Secondary:

8. Keywords for Experimental System/Species:

• Species:
• Study Type:
  • In Vitro
  • In Vivo
  • Laboratory Study
• Fate and Transport:

9. Keywords for Experimental Endpoints:

• Health Effect:
• Hormonal Measures:
  • Peptide Hormones
• Level Of Study:
  • Gene Expression
• Chemistry Metabolism:
  • Tissue Residue
  • Ah Receptor
• Life Stage:
• Risk Assessment:

10. Chemical Agents:

• Dioxins
• Pahs

11. Performing Institution:

• UNIVERSITY OF WISCONSIN MADISON

12. Contact:

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