Centrosome are involved in the organization of cytoarchitecture in cells from yeast to man. As organizers of microtubules (microtubule organizing centers, MTOCs), centrosomes are involved in numerous fundamental cell processes including cell migration and cell division. Thus, and understanding of the composition and biochemical regulation of centrosomes will have an impact on understanding cancer, metastasis, chemotaxis, and aspects of early development. Methods have been developed to take advantage of the unique characteristics of clam oocytes which allow the purification of sufficient quantities of centrosomes for preparative and analytical biochemistry. The experiments outlined in this proposal will define the protein composition of centrosomes and determine if compositional changes are a function of the meiotic cell cycle. Functional in vitro assays coupled with video- and electron-microscopy will be used to quantitate centriole content and the ability of purified centrosomes to direct microtubule polymerization. Monoclonal antibodies will be generated for the identification and purification of molecules (MTOCs) which direct microtubule organization. Finally, key experiments will be carried out to determine if centrosomes can be induced to mature in vitro, as indicated by changes in their ability to form and replicate centrioles or their ability to organize microtubules. These studies will open the way for classic biochemical approaches designed to purify molecules which regulate centrosomes during the cell cycle.
| Shang, Wen; Crone, Donna E; Yang, Hoichang et al. (2009) Using centrosome fragments in the directed assembly of microtubules. J Nanosci Nanotechnol 9:871-5 |
| DiMaio, Michael A; Mikhailov, Alexei; Rieder, Conly L et al. (2009) The small organic compound HMN-176 delays satisfaction of the spindle assembly checkpoint by inhibiting centrosome-dependent microtubule nucleation. Mol Cancer Ther 8:592-601 |
| Sankaran, Satish; Crone, Donna E; Palazzo, Robert E et al. (2007) Aurora-A kinase regulates breast cancer associated gene 1 inhibition of centrosome-dependent microtubule nucleation. Cancer Res 67:11186-94 |
| Sankaran, Satish; Crone, Donna E; Palazzo, Robert E et al. (2007) BRCA1 regulates gamma-tubulin binding to centrosomes. Cancer Biol Ther 6:1853-7 |
| Shang, Wen; Dordick, Jonathan S; Palazzo, Robert E et al. (2006) Direct patterning of centrosome arrays as templates for the assembly of microtubules. Biotechnol Bioeng 94:1012-6 |
| Alliegro, Mark C; Alliegro, Mary Anne; Palazzo, Robert E (2006) Centrosome-associated RNA in surf clam oocytes. Proc Natl Acad Sci U S A 103:9034-8 |
| Schnackenberg, B J; Palazzo, R E (2001) Reconstitution of centrosome microtubule nucleation in Spisula. Methods Cell Biol 67:149-65 |
| Nilsson, H; Steffen, W; Palazzo, R E (2001) In vitro reconstitution of fish melanophore pigment aggregation. Cell Motil Cytoskeleton 48:1-10 |
| Schnackenberg, B J; Hull, D R; Balczon, R D et al. (2000) Reconstitution of microtubule nucleation potential in centrosomes isolated from Spisula solidissima oocytes. J Cell Sci 113 ( Pt 6):943-53 |
| Palazzo, R E; Vogel, J M; Schnackenberg, B J et al. (2000) Centrosome maturation. Curr Top Dev Biol 49:449-70 |
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