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Related Experiment Videos

Isolation of a cDNA clone encoding a biologically active thyroid hormone receptor.

R J Koenig1, R L Warne, G A Brent

  • 1Howard Hughes Medical Institute Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.

Proceedings of the National Academy of Sciences of the United States of America
|July 1, 1988
PubMed
Summary

Researchers identified a novel beta-type c-erbA protein (erb62) that functions as a biologically active thyroid hormone (T3) receptor. This discovery advances understanding of T3 receptor mechanisms in target tissues.

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Area of Science:

  • Molecular Biology
  • Endocrinology
  • Genetics

Background:

  • The c-erbA gene family encodes proteins that function as thyroid hormone receptors.
  • Thyroid hormones regulate numerous physiological processes, including metabolism and development.
  • Understanding the specific functions and interactions of c-erbA subtypes is crucial for deciphering thyroid hormone signaling.

Purpose of the Study:

  • To isolate and characterize a novel c-erbA cDNA clone from GH3 cells.
  • To determine if the encoded protein exhibits thyroid hormone binding and biological activity.
  • To investigate the tissue distribution and functional role of the identified c-erbA protein.

Main Methods:

  • Isolation of a c-erbA cDNA clone (erb62) from a GH3 cell library.

Related Experiment Videos

  • Characterization of the encoded protein's size and amino acid sequence.
  • Binding assays to assess affinity for 3,5,3'-triiodothyronine (T3) and its analogs.
  • RNA gel blot analysis to determine tissue-specific expression.
  • COS-cell transient cotransfection assays to evaluate biological activity using a reporter gene system.

    • Main Results:

    • A 4.5-kilobase cDNA clone, erb62, encoding a 461-amino acid beta-type c-erbA protein was isolated.
    • The erb62 protein demonstrated high-affinity binding to T3 and its analogs, comparable to authentic T3 receptors.
    • RNA analysis revealed hybridization to a 6-kilobase RNA in organs known to express T3 receptors, including heart, kidney, and brain.
    • Cotransfection experiments confirmed that erb62 encodes a biologically active T3 receptor capable of mediating T3-inducible gene expression.

    Conclusions:

    • The isolated erb62 clone represents a functional beta-type c-erbA protein that acts as a T3 receptor.
    • This protein is expressed in key target organs, suggesting a significant role in thyroid hormone action.
    • The findings contribute to the understanding of thyroid hormone receptor diversity and function.