The stereochemistry and thermodynamics of DNA-target recognition by specific DNA-binding proteins will be studied. The model system used is the E2 transcriptional regulatory protein and replication factor of the cancer- associated papillomaviruses. The techniques used will include X-ray crystallography, titration calorimetry, quantitative gel-retardation assays and mutational analyses of the proteins. The following specific goals will be pursued: To elucidate the structural basis for DNA target discrimination by the E2 protein of the human papillomaviruses. To establish the energetics of specific DNA recognition by the E2 proteins. To determine the role of the 'hinge' region of the E2 repressor proteins in DNA-binding. To determine the three-dimensional structure of the entire B2 protein and its complex with DNA. This study will go beyond the cataloguing of specifying intermolecular contacts. It will explicitly define and quantitate the contributions of indirect factors such as intrinsic deformability and changes in solvent accessible surface areas in mediating the DNA target selectivity exhibited by the E2 proteins. Insights regarding the contributions to specificity and affinity of these features will then be extrapolated to other protein-DNA systems. The detailed structural information will provide a basis for the rational design of molecules that could modulate E2-DNA complex formation. Such agents would be of therapeutic value in the treatment of papillomavirus-caused pre-cancerous lesions.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA066964-04
Application #
2871852
Study Section
Virology Study Section (VR)
Program Officer
Wong, May
Project Start
1996-02-21
Project End
2001-01-31
Budget Start
1999-02-01
Budget End
2001-01-31
Support Year
4
Fiscal Year
1999
Total Cost
Indirect Cost
Name
New York University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10016
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