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The beta-globin stage selector element factor is erythroid-specific promoter/enhancer binding protein NF-E4
J L Gallarda1, K P Foley, Z Y Yang
1Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208.
Genes & Development
|December 1, 1989
Summary
Researchers identified NF-E4, a protein crucial for adult beta-globin gene activation in chicken red blood cells. This discovery offers a new method for studying scarce transcriptional regulatory proteins.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Analyzing scarce transcriptional regulatory proteins is challenging due to low abundance.
- Traditional biochemical purification methods are labor-intensive and costly.
Purpose of the Study:
- To develop an alternative strategy for identifying and characterizing trans-acting factors.
- To analyze proteins interacting with the chicken adult beta-globin gene enhancer and promoter.
Main Methods:
- Utilized a sensitive 'reverse' radioimmunoassay to identify antibodies against sequence-specific DNA-binding proteins.
- Employed a monoclonal antibody-based DNase I footprint selection technique to pinpoint specific DNA-binding proteins.
Main Results:
- Identified a 65-kD protein, named NF-E4, exclusively present in mature definitive (adult) chicken erythroid cells.
- Demonstrated that NF-E4 binds to sequences in both the beta-globin promoter and enhancer.
- Showed NF-E4 is the trans-acting factor responsible for adult beta-globin gene stage-specific activation.
Conclusions:
- The developed methodology is broadly applicable for studying any trans-acting regulatory factor with an identifiable DNA-binding sequence.
- NF-E4 plays a critical role in conferring definitive erythrocyte stage-specific transcriptional activation to the adult beta-globin gene.