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Adenovirus DNA binding protein: helix destabilising properties
A Monaghan1, A Webster, R T Hay
1School of Biological and Medical Sciences, University of St Andrews, Fife, UK.
Nucleic Acids Research
|March 11, 1994
Summary
Adenovirus DNA binding protein unwinds DNA duplexes without ATP, crucial for viral replication. Its C-terminal fragment contains this essential DNA unwinding activity.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Adenovirus DNA binding protein is vital for viral DNA replication.
- Its precise role in DNA replication requires further investigation.
Purpose of the Study:
- To investigate the DNA binding protein's interaction with DNA duplexes.
- To elucidate the protein's DNA unwinding mechanism and functional domains.
Main Methods:
- In vitro DNA unwinding assays using M13 phage DNA and synthetic DNA duplexes.
- Investigating the effects of DNA stability, G+C content, and transcription factor binding.
- Protein cleavage and domain mapping using chymotrypsin and monoclonal antibodies.
Main Results:
- Adenovirus DNA binding protein efficiently unwound partial DNA duplexes without ATP or MgCl2.
- Unwinding activity was sensitive to DNA duplex stability and G+C content for double-stranded DNA.
- Transcription factor binding at specific sites inhibited protein-mediated unwinding.
- The C-terminal fragment (39,000 MW) of the protein retained DNA unwinding activity.
Conclusions:
- The DNA binding protein's unwinding activity is essential for viral DNA replication processes.
- This activity, located in the C-terminal domain, likely destabilizes DNA duplexes during replication initiation and elongation.