Centriole assembly regulation by Plk4
Abstract/Contents
- Abstract
- Regulation of centriole number is important to ensure proper cell division and formation of the correct number of cilia, a signaling organelle. Activity of the polo-like kinase Plk4 regulates the number of centrioles formed each cell cycle. To study Plk4 function and the mechanism of centriole assembly we developed a system in Xenopus laevis egg extract, where we show Plx4 (Xenopus Plk4) overexpression drives de novo centriole assembly. Centriole assembly regulation in this system mimicked regulation of centriole duplication in somatic cells, suggesting that mechanisms identified in the egg extract would apply to centriole formation in general. By analyzing the functionality of Plx4 mutants we determined that Plx4 activity is not regulated by proteolysis in vitro in the extract, and likely not in vivo during early embryogenesis. To identify potential Plk4 regulators and effectors we took two approaches in egg extract: 1) chemical genetics to specifically modify Plk4 substrates in egg extract, and 2) pull-down and mass spectrometry identification of co-purifying proteins. Using the second approach we identified an interaction between Plx4 and the centrosomal protein Cep152. We found that Cep152 directly interacts with human Plk4, and that Cep152 is required for centriole duplication and Plk4-based centriole amplification. Cep152 is required to recruit the early centriole assembly protein Sas-6, but not Plk4, to the centriole. Finally, Cep152 was phosphorylated by Plk4 in vitro, suggesting that Cep152 acts with Plk4 to initiate centriole formation.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Copyright date | 2011 |
| Publication date | 2010, c2011; 2010 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Hatch, Emily Marian |
|---|---|
| Associated with | Stanford University, Department of Biology |
| Primary advisor | Stearns, Tim |
| Thesis advisor | Stearns, Tim |
| Thesis advisor | Cyert, Martha S, 1958- |
| Thesis advisor | Ferrell, James Ellsworth |
| Thesis advisor | Nelson, W. J. (W. James) |
| Advisor | Cyert, Martha S, 1958- |
| Advisor | Ferrell, James Ellsworth |
| Advisor | Nelson, W. J. (W. James) |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Emily Hatch. |
|---|---|
| Note | Submitted to the Department of Biology. |
| Thesis | Ph.D. Stanford University 2011 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2011 by Emily Marian Hatch
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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