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A universal target sequence is bound in vitro by diverse homeodomains
1Department of Biochemistry and Biophysics, University of California at San Francisco 94143-0448.
Mechanisms of Development
|September 1, 1993
Summary
Researchers identified 20 DNA binding domains, including 17 homeodomains, that bind a specific DNA sequence. This study reveals DNA binding specificity is more conserved than amino acid sequence in homeodomain evolution.
Area of Science:
- Molecular Biology
- Genetics
- Evolutionary Biology
Background:
- The Engrailed gene plays a crucial role in embryonic development.
- Understanding DNA binding proteins is essential for deciphering gene regulation.
Purpose of the Study:
- To identify DNA binding proteins that recognize the Engrailed binding site.
- To investigate the evolutionary conservation of DNA binding specificity and amino acid sequences in homeodomains.
Main Methods:
- Screening a Drosophila cDNA library using a consensus Engrailed binding site.
- Bacteriophage expression vector utilized for cloning and expression.
- Oligonucleotide binding assays to confirm sequence specificity.
Main Results:
- Identified 20 distinct DNA binding domains, with 17 being homeodomains.
- Confirmed related sequence specificity for the retrieved binding domains.
- Observed remarkable diversity in amino acid sequences among the identified homeodomains.
Conclusions:
- DNA binding specificity is highly conserved during homeodomain evolution.
- Amino acid sequence divergence does not necessarily correlate with changes in DNA binding specificity.