The Structural Cell Biology (SCB) Core provides critical technologies and support for the Structural CellBiology of DNA Repair Machines (SBDR) Program. Major challenges of SBDR stem from the dynamic andcoordinated assembly of large protein complexes involved in DNA repair processes. These complexesundergo functionally important conformational changes and modifications. The SCB Core will providestructural expertise and technologies suitable for SBDR project and program Aims, create a functional bridgebetween atomic resolution structures and molecular envelopes, and help close the gap between static crystalstructures and biologically relevant, multi-component macromolecular machines. In particular, the SCB Corewill provide SBDR with three distinct and complementary methods for structural analyses. (1) Multiwavelengthsingle crystal X-ray diffraction will provide high-resolution structures of discrete states. (2) SmallAngle X-ray Scattering (SAXS) will characterize the solution dynamics of protein complexes by visualizingflexible regions and induced conformational changes. (3) Scanning Force Microscopy (SFM) of singlemolecules, available through the Wyman lab, will provide information about DNA and protein dynamics, andwill reveal structural insight into heterogeneous mixtures previously inaccessible using crystallographic orSAXS techniques. The SCB Core is designed to supply the SBDR projects with the necessary tools andproficiency to overcome the structural biology challenges inherent to analysis of large complexes. Therequested funding provides staff to maximize interaction with the EMB Core and for SBDR use of theStructurally Integrated Biology for Life Sciences (SIBYLS) beamline at the Advanced Light Source (ALS) atthe Lawrence Berkeley National Laboratories (LBNL). The SIBYLS beamline is a unique synchrotronresource that provides tunable wavelengths for both single crystal X-ray diffraction and SAXS. The SCBCore will develop software that addresses current limitations in the analysis of DNA repair proteins includingsoftware that will combine results from high and low resolution techniques through the systematic andobjective fitting of X-ray crystal structures into molecular envelopes generated by EM and SAXSexperiments. The SCB Core will test, develop, and provide advanced tools to detect and measure posttranslationalmodifications. Understanding the dynamic structures of macromolecular machines for DNArepair will generate insights into multi-component systems that have remained elusive through the study ofindividual component biomolecules. The results from the SCB Core will be applied to the understanding ofcancer etiology and potential cancer diagnostics and prognostics through interactions with the UCSFComprehensive Cancer Center.
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