The overall objective of our research program is to decipher the structure-function relationship for the human adenovirus DNA binding protei (DBP) and define the mechanistically how this protein performs several of its different functions. DBP is the paradigm for eukaryotic ssDNA binding proteins. Thus insights into its functions and how they are dictated by its structure will serve as an important model for this class of proteins. We will continue to use our genetic approach to study several aspects of DBP structure and function. The role(s) of phosphorylation in auto- enhancement and possibly in nuclear localization will be defined. The contributions made to nucleic acid binding and DNA replication by both DBP's presumptive metal binding finger and aa's in its DNA binding site will be determined. In addition ne mutations, particularly in the segment of the N-t domain which is responsible for regulating late gene expression and the virus' host range, will be isolated and characterized. The genetic approach is being combined with the biochemical characterization of the mutant-infected (transfected) cells or the mutant DBPs. For example, studies are proposed to determine whether DBP enhances its expression at the transcriptional level and if so via which promotor (E2aE or E2aL) and promotor element(s)? The region (aa's) of the protein involved in ssDNA binding is also being physically mapped; and proteins associated with DBP will be identified and characterized.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023591-10
Application #
2062287
Study Section
Virology Study Section (VR)
Project Start
1990-09-01
Project End
1996-07-31
Budget Start
1994-08-01
Budget End
1996-07-31
Support Year
10
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Rutgers University
Department
Type
Organized Research Units
DUNS #
038633251
City
New Brunswick
State
NJ
Country
United States
Zip Code
08901
Ricigliano, J W; Brough, D E; Klessig, D F (1994) Identification of a high-molecular-weight cellular protein complex containing the adenovirus DNA binding protein. Virology 202:715-23
Cleghon, V; Piderit, A; Brough, D E et al. (1993) Phosphorylation of the adenovirus DNA-binding protein and epitope mapping of monoclonal antibodies against it. Virology 197:564-75
Brough, D E; Droguett, G; Horwitz, M S et al. (1993) Multiple functions of the adenovirus DNA-binding protein are required for efficient viral DNA synthesis. Virology 196:269-81
Carter, B J; Antoni, B A; Klessig, D F (1992) Adenovirus containing a deletion of the early region 2A gene allows growth of adeno-associated virus with decreased efficiency. Virology 191:473-6
Eagle, P A; Klessig, D F (1992) A zinc-binding motif located between amino acids 273 and 286 in the adenovirus DNA-binding protein is necessary for ssDNA binding. Virology 187:777-87
Brough, D E; Cleghon, V; Klessig, D F (1992) Construction, characterization, and utilization of cell lines which inducibly express the adenovirus DNA-binding protein. Virology 190:624-34
Cleghon, V; Klessig, D F (1992) Characterization of the nucleic acid binding region of adenovirus DNA binding protein by partial proteolysis and photochemical cross-linking. J Biol Chem 267:17872-81
Vos, H L; Brough, D E; Van der Lee, F M et al. (1989) Characterization of adenovirus type 5 insertion and deletion mutants encoding altered DNA binding proteins. Virology 172:634-42
Silverman, L; Cleghon, V; Klessig, D F (1989) Increased permissivity of monkey cells to human adenovirus multiplication is affected by culturing conditions and correlates with both synthesis of virion fiber protein and altered splicing of its mRNA. Virology 173:109-19
Silverman, L; Klessig, D F (1989) Characterization of the translational defect to fiber synthesis in monkey cells abortively infected with human adenovirus: role of ancillary leaders. J Virol 63:4376-85

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