BIOCHEMICAL CHARACTERIZATION OF BACTERIAL CYTOKINESIS

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DESCRIPTION (provided by applicant): Cell division will be biochemically analyzed in the filamentous prokaryote, Streptomyces coelicolor. This species is advantageous for this study because growth and viability do not require cell division, a collection of division null mutants already exists, and reagents vital to the success of this project have been prepared. Mounting evidence suggests that the division proteins function as a complex that is assembled in an ordered fashion; however direct biochemical support for the complex is absent. Many division proteins contain at east one membrane-spanning segment, making the biochemical characterization of the complex more challenging. Therefore, the time has come to apply membrane protein biochemical techniques, which will require training in the area. Specifically, I will identify the proteins in the Streptomyces cell division complex using three unique but complementary biochemical attacks: cross-linking, immune precipitation, and column Chromatography. These techniques have successfully defined other integral membrane protein machines. I also hope to identify novel components of the cell division complex using proteomics. The net result of this project will uniquely equip my laboratory to continue to make significant strides in the cell division field.