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FP7

CENTRIOLE_LENGTH Résumé de rapport

Project ID: 623275
Financé au titre de: FP7-PEOPLE
Pays: Switzerland

Final Report Summary - CENTRIOLE_LENGTH (Molecular basis of centriole length control)

Summary description of the project objectives

Centrioles play a key role in the assembly of the centrosome, the major microtubule organizing centre, and are crucial for the formation of cilia and flagella [1]. Significant progress has been made toward understanding centriole biogenesis, but the mechanisms that determine centriole length and prevent centrioles from forming cilia or flagella remain unknown. CP110, Cep97, Cep120, Cep104, CPAP, SPICE1 and centrobin have been identified as key players in these fundamental biological processes [2-6]. The overarching aim of my project is to understand how these proteins cooperate in order to control centriole length. The following unresolved questions are being addressed:

1) What is the molecular mechanism of the interactions between centriolar proteins that play a key role in controlling centriole length and cilia formation?

2) How do the proteins involved in centriole length control interact with tubulin and microtubules?

3) What is the functional importance of interactions of key proteins in controlling centriole length and cilia formation?

A multidisciplinary approach encompassing state-of-the-art bioinformatics, biochemical, biophysical, and structural and cell biology methods is used to tackle these open questions. The results will significantly contribute to our understanding of centriole and cilia biogenesis. Ultimately, the proposed research will help to understand how misregulation of these processes leads to human diseases such as cancer, polycystic kidney disease, microcephaly, dwarfism, and primary ciliary dyskinesia [7].

Summary overview of results obtained during the project

- Discovery that Cep104 is a novel tubulin binding protein
- Crystal structure of the Cep104 TOG domain that provided a rational explanation on the interaction between the TOG domain and tubulin
- Biophysical/biochemical/functional characterization of Cep104/Cep97, Cep104/CP110 and Cep104/tubulin complexes
- Publication describing the Cep104 interaction network prepared for submission
- Biophysical/biochemical/functional characterization of p60-MIT:p80-CTD, p60-MIT:p80-CTD:CAMSAP and p60-MIT:p80-CTD:ASPM complexes
- Crystal structures of p60-MIT:p80-CTD, p60-MIT:p80-CTD:CAMSAP and p60-MIT:p80-CTD:ASPM complexes
- First draft of a publication describing the structures and functions of p60-MIT:p80-CTD and p60-MIT:p80-CTD:ASPM complexes
- Accepted open access publication describing the membrane targeting and activation of human PI4KB by the ACBD3 protein to which I have contributed during the second year [8]
- Accepted open access publication describing the multi-lab AUC reference study to which I contributed during the first year [9]

Conclusions and the socio-economic impacts of the project

- biochemical, biophysical, structural, and functional information about Cep104:Cep97,
Cep104:CP110 and Cep104:tubulin, that was uncovered during this study provides a solid platform for performing further experiments with the aim to understand centriole length control and cilliogenesis
- deciphering the process of centriole length control will significantly contribute to our understanding
of human ciliary diseases, such as renal diseases and polycystic kidney disease
- biochemical, biophysical, structural, and functional information about the microtubule severing enzyme katanin offered new insights into the process of microtubule severing
- as katanin has been recently identified as one of the proteins responsible for microcephaly [10, 11], the biochemical, biophysical, structural, and functional information will contribute to our understanding of this disease
- biochemical, biophysical, structural, and functional information about the katanin/CAMSAP and katanin/ASPM complexes revealed their major role in regulating microtubule minus end dynamics
- understanding microtubule dynamics is crucial as microtubules are targeted by many anticancer drugs
- all the results were/will be presented in the form of high impact publications and in the form of oral and/or poster presentations at international conferences
- once the results are accepted for publications the important results will be presented to the public in a general and comprehensive way in the form of press releases.

References

1. Gonczy, P. (2012) Towards a molecular architecture of centriole assembly, Nature reviews Molecular cell biology. 13, 425-35.
2. Spektor, A., Tsang, W. Y., Khoo, D. & Dynlacht, B. D. (2007) Cep97 and CP110 suppress a cilia assembly program, Cell. 130, 678-90.
3. Comartin, D., Gupta, G. D., Fussner, E., Coyaud, E., Hasegan, M., Archinti, M., Cheung, S. W., Pinchev, D., Lawo, S., Raught, B., Bazett-Jones, D. P., Luders, J. & Pelletier, L. (2013) CEP120 and SPICE1 Cooperate with CPAP in Centriole Elongation, Current biology : CB. 23, 1360-6.
4. Gudi, R., Zou, C., Li, J. & Gao, Q. (2011) Centrobin-tubulin interaction is required for centriole elongation and stability, The Journal of cell biology. 193, 711-25.
5. Schmidt, T. I., Kleylein-Sohn, J., Westendorf, J., Le Clech, M., Lavoie, S. B., Stierhof, Y. D. & Nigg, E. A. (2009) Control of centriole length by CPAP and CP110, Current biology : CB. 19, 1005-11.
6. Satish Tammana, T.V., Tammana, D., Diener, D.R., Rosenbaum, J. (2013) Centrosomal protein 5018-29
7. Nigg, E. A. & Raff, J. W. (2009) Centrioles, centrosomes, and cilia in health and disease, Cell. 139, 663-78.
8. Klima, M., Tóth, DJ., Hexnerova, R., Baumlova, A., Chalupska, D., Tykvart, J., Rezabkova, L., Sengupta, N., Man, P.,5, Dubankova, A., Humpolickova, J., Nencka, R., Veverka, V., Balla, T. & Boura, E. (2016) Structural insights and in vitro reconstitution of membrane targeting and activation of human PI4KB by the ACBD3 protein. Sci Rep. 6, 23641.
9. Zhao, H., Ghirlando, R., ... Rezabkova, L., et al. (2015) A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation. PLoS One, 10(5):e0126420
10. Hu, W.F., Pomp, O., Ben-Omran, T., et al., Neuron, 84, 1240-57.
11. Mishra-Gorur, K., Ça#layan, A.O., Schaffer, A.E., et al. (2014) Mutations in KATNB1 cause complex cerebral
malformations by disrupting asymmetrically dividing neural progenitors. Neuron, 84, 1226-39.

Informations connexes

Résultat en bref

Contact

Irene Walthert
Tél.: +41 56 310 2664
E-mail
Numéro d'enregistrement: 184429 / Dernière mise à jour le: 2016-06-24