The central theme of the proposal is to analyze how initiator proteins activate replication origins by """"""""action at a distance"""""""". Specifically, long range activation of distant origins by pi mediated enhancer-origin interaction, shutting down of the potential origin at the enhancer of plasmid R6K will be analyzed by a multifaceted approach, structure-function analysis of normal pi and mutants by both molecular genetic and structural analysis by X-ray crystallography and NMR spectroscopy. The shorter range looping of pSC101 origin promoted by DnaA protein and the role of RepA and the DNA-binding protein IHF will also be investigated. Finally, the """"""""handcuffing"""""""", i.e. protein mediated intermolecular pairing of origin interon as a viable model for copy control will be critically examined in vivo and in vitro by isolation and analysis of mutants defective in the process. The investigators will also analyze whether other origin-binding proteins modulate and perhaps enhance or reverse handcuffing process.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37AI019881-16
Application #
2503937
Study Section
Special Emphasis Panel (NSS)
Project Start
1983-07-01
Project End
2003-06-30
Budget Start
1998-07-01
Budget End
1999-06-30
Support Year
16
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Duke University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705
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