Related Experiment Videos
Synergy between adjacent zinc fingers in sequence-specific DNA recognition
1Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom.
Summary
Zinc fingers, DNA-binding proteins, can recognize overlapping DNA sequences. This interaction between adjacent fingers enhances sequence specificity, impacting novel zinc finger design.
Area of Science:
- Molecular Biology
- Structural Biology
- Biochemistry
Background:
- Zinc fingers are modular DNA-binding proteins.
- Typically, each finger recognizes three base pairs in discrete subsites.
- Crystallographic data suggested potential inter-finger contacts.
Purpose of the Study:
- To investigate the DNA binding specificity of the Zif268 protein.
- To determine if inter-finger contacts influence binding specificity.
- To explore implications for designing novel zinc finger proteins.
Main Methods:
- Studied the three-finger DNA-binding domain of Zif268.
- Utilized a panel of Zif268 variants.
- Assessed effects of deleting a putative inter-finger contact on DNA binding specificity.
Main Results:
- Deleting a contact in finger 3 altered specificity for the 5' base of finger 2's subsite.
- Demonstrated that Zif268-like fingers can specify overlapping 4-base pair subsites.
- Showed that synergy between adjacent fingers creates boundary sequence specificity.
Conclusions:
- Zif268-like zinc fingers can recognize overlapping DNA subsites.
- Inter-finger synergy is crucial for sequence specificity at subsite boundaries.
- Findings inform the design of zinc fingers with tailored DNA-binding properties.