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A cellular protein binds to a conserved sequence in the adenovirus type 2 enhancer
Nucleic Acids Research
|March 25, 1987
Summary
Researchers identified a specific protein factor that binds to critical DNA sequences in adenovirus type 2 enhancers. This enhancer binding factor also interacts with similar sequences in SV40 enhancers, suggesting a common mechanism for viral gene regulation.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Enhancers are crucial regulatory DNA elements that control gene transcription.
- Understanding enhancer-binding proteins is key to deciphering viral and cellular gene expression.
- Adenovirus type 2 and Simian Virus 40 (SV40) enhancers share conserved sequences.
Purpose of the Study:
- To identify and characterize proteins that bind to the adenovirus type 2 enhancer.
- To investigate the DNA binding specificity and affinity of these enhancer-binding factors.
- To determine if these factors interact with conserved sequences common to viral enhancers.
Main Methods:
- Gel retention assay to detect protein-DNA interactions.
- Chromatography (DEAE-Sepharose and heparin-Sepharose) for protein purification.
- DNase protection assays to map DNA binding sites.
- Competition experiments using synthetic oligonucleotides.
Main Results:
- Three distinct enhancer binding factors were resolved from HeLa nuclear extracts.
- A purified factor predominantly binds to the conserved sequence GTGGAAATTT (positions 144-181) in the adenovirus type 2 enhancer.
- This factor also binds to homologous sequences in the adenovirus Ela upstream region and exhibits high-affinity binding to the SV40 enhancer.
Conclusions:
- A specific enhancer binding factor interacts with a conserved DNA motif present in adenovirus type 2 and SV40 enhancers.
- This factor plays a role in regulating gene expression through common enhancer sequences.
- The findings suggest shared mechanisms of transcriptional regulation by viral enhancers.