Core B is a synthesis, analysis and biochemical core that will provide indispensable services for all three projects. The synthetic component of Core B will carry out scaled-up synthesis, purification and analysis of "tool-kit" and other nucleotide analogs for use in Projects 1 and 3. The biochemical component of Core B is designed to carry out protein purifications and to synthesize specialized DNA constructs required for Projects 1 and 3. The analysis component will be responsible for acquiring and performing global analysis on kinetic data, including presteady state stopped-flow fluorescence intensity and rotational anisotropy, rapid quench experiments, as well as basic NMR and EPR-ENDOR analysis, to be used in support of Projects 2 and 3. Core B will also serve as a primary vehicle for training graduate and postdoctoral students in chemical synthesis and protein purification techniques, the principles and practice of enzyme kinetic analysis, and techniques in experimental NMR and EPR spectroscopy.
Core B provides essential chemical probes, biochemical assays and spectroscopic analysis in support of the three component projects of this Program Project. This work will lead to better understanding of the structure, fidelity and catalytic mechanism of DNA polymerases, and also includes synthesis of compounds to develop a new approach to selective inhibition of cancer-related DNA repair by DNA polymerase beta.
|Kirby, Thomas W; Derose, Eugene F; Beard, William A et al. (2014) Substrate rescue of DNA polymerase ? containing a catastrophic L22P mutation. Biochemistry 53:2413-22|
|Wu, Sangwook; Beard, William A; Pedersen, Lee G et al. (2014) Structural comparison of DNA polymerase architecture suggests a nucleotide gateway to the polymerase active site. Chem Rev 114:2759-74|
|Prasad, Rajendra; Horton, Julie K; Chastain 2nd, Paul D et al. (2014) Suicidal cross-linking of PARP-1 to AP site intermediates in cells undergoing base excision repair. Nucleic Acids Res 42:6337-51|
|Freudenthal, Bret D; Beard, William A; Wilson, Samuel H (2014) Watching a DNA polymerase in action. Cell Cycle 13:691-2|
|Towle-Weicksel, Jamie B; Dalal, Shibani; Sohl, Christal D et al. (2014) Fluorescence resonance energy transfer studies of DNA polymerase ?: the critical role of fingers domain movements and a novel non-covalent step during nucleotide selection. J Biol Chem 289:16541-50|
|Beard, William A; Wilson, Samuel H (2014) Structure and mechanism of DNA polymerase ?. Biochemistry 53:2768-80|
|Oertell, Keriann; Chamberlain, Brian T; Wu, Yue et al. (2014) Transition state in DNA polymerase ? catalysis: rate-limiting chemistry altered by base-pair configuration. Biochemistry 53:1842-8|
|Hwang, Candy S; Kashemirov, Boris A; McKenna, Charles E (2014) On the observation of discrete fluorine NMR spectra for uridine 5'-?,?-fluoromethylenetriphosphate diastereomers at basic pH. J Org Chem 79:5315-9|
|Seamon, Kyle J; Hansen, Erik C; Kadina, Anastasia P et al. (2014) Small molecule inhibition of SAMHD1 dNTPase by tetramer destabilization. J Am Chem Soc 136:9822-5|
|Sassa, Akira; Ça?layan, Melike; Dyrkheeva, Nadezhda S et al. (2014) Base excision repair of tandem modifications in a methylated CpG dinucleotide. J Biol Chem 289:13996-4008|