Cycle cellulaire et développement
LIONEL PINTARD
The Cell Cycle and Development team aims to acquire new knowledge to decipher the mechanisms of cell division to understand the mechanisms of cancer, a disease resulting from uncontrolled cell division.
The team mainly uses the nematode C. elegans as a model system and employs a multidisciplinary approach combining various approaches (biochemistry, genetics, imaging, proteomics) to ask questions at different scales, from the molecule to the organism. As the mechanisms of regulation of cell division are conserved between species, the team is also studying the emerging paradigms in C. elegans in human cells.
Keywords: Cell division – Mitosis – Meiosis –Kinases – Microtubule-severing enzymes – Nuclear envelope breakdown
+33 (0)1 57 27 80 89 Contact @LabPintard https://sites.google.com/site/pintardlab/
Our vision
Acquiring new knowledge to decipher how cells divide, which is key to better understand the mechanisms of cancer, a disease caused by uncontrolled cell division.
Context
Humans are roughly built from 1013 cells corresponding to 200 different cell types. All these cells are generated through cell divisions, starting from a single cell, the fertilized egg. To generate these large number of cells and maintain tissue homeostasis, the human body experiences as many as 1016 cell divisions in a lifetime. During each cell division, the genome must be faithfully replicated and equally segregated between the daughter cells during mitosis. Defects in these processes can have drastic consequences, leading to outcomes such as cell death, genome instability or deregulated growth, typical of cancer. Despite considerable progress, the mechanisms regulating cell division are incompletely understood. This lack of knowledge has considerably limited the development of innovative therapeutic approaches.
Research programm
I- Mechanisms regulating mitotic entry in space and time
Commitment to mitosis must be tightly coordinated with DNA replication to preserve genome integrity. Consistently, unscheduled mitosis may contribute to genetic instability. Entry into mitosis is controlled by evolutionarily conserved serine/threonine kinases (Aurora A, Polo-like kinase, Plk1) as well as counteracting phosphatases (PPases). How these kinase activities are regulated in space and time, and how they work in concert to trigger mitosis at the right time remain ill defined. In this context, we are investigating the
- Activation mechanism of key mitotic kinases (Aurora A, Polo-like kinase 1)
- Role of mitotic kinases in nuclear envelope breakdown (NEBD)
- Role and regulation of Mitotic kinases during asynchronous cell division
Figure 1: Bora-Aurora A (Aurka)-Plk1 axis during mitotic entry
II- Role and regulation of the microtubule-severing enzyme Katanin
Microtubules (MTs) are dynamic cytoskeleton polymers, which play a central role in cell division, morphogenesis, motility and signaling. Most MT regulatory proteins interact with the plus or minus end of microtubules and thereby control their polymerization and depolymerization rates. However, one class of MT regulator interacts with the MT lattice and severs MTs throughout their length, thereby controlling their size and density in the cell. Three evolutionarily conserved MT-severing enzymes have been identified: Fidgetin, Spastin and Katanin. Mutations of these enzymes have been linked to various defects and pathologies including developmental defects and neurodegenerative disorders. However, little is known about the precise mechanisms by which these enzymes sever MTs. Likewise, how these enzymes are regulated in space and time is poorly understood. We are currently focusing on deciphering the mode of action and regulation of Katanin, which is essential for female meiotic spindle assembly in C. elegans. We are focusing our work on the
- role of MT-severing in meiotic spindle assembly
- Mechanism of MT-severing by Katanin
- Regulation of Katanin-mediated MT-severing in space and time during development
Figure 2: Role and regulation of Katanin during C. elegans development (From Joly et al. JCB 2020).
III- Cullin-RING E3-Ligases in cell division
Cullin-RING E3-ligases (CRL) represent the largest family of E3 ubiquitin-ligases targeting the degradation of key cell cycle regulators in space and time, contributing to the orderly progression of the cell division cycle. We are interested in understanding how these enzymes regulate cell cycle progression in a developmental context.
- CRL in the regulation of the Bora-Aurora-Plk1 pathway
- CRL in the regulation of Katanin activity
- CRL in the maintenance of DNA replication integrity
Methods
We use a multidisciplinary approach including biochemistry (reconstitution of enzymatic activities from purified components to dissect molecular mechanisms), genetics, live cell imaging, proteomics approaches using both human cells and the nematode C. elegans. Mechanisms regulating cell division are conserved between species such that emerging paradigm from C. elegans can be immediately investigated in human cells. Furthermore, C. elegans offers a number of practical advantages to study conserved pathways regulating cell division (Pintard & Bowerman, Wormbook, Genetics 2019).
Members
To contact a member of the team by e-mail: name.surname@ijm.fr
June, 2021 (c) Pintard Lab
From left to right: Anais, Griselda, Batool, Lucie, NicoT, Sylvia, Lionel, Eva, Lola, Anaelle, Emma, NicoJ
Velez-Aguilera, G., Ossareh-Nazari, B., Van Hove, L., Joly, N., and Pintard, L*. (2022). Cortical microtubule pulling forces contribute to the union of the parental genomes in the. Elife Mar 8;11:e75382. doi: 10.7554/eLife.75382.
Tavernier, N., Thomas, Y., Vigneron, S., Maisonneuve, P., Orlicky, S., Mader, P., Regmi, S. G., Van Hove, L., Levinson, N. M., Gasmi-Seabrook, G., Joly, N., Poteau, M., Velez-Aguilera, G., Gavet, O., Castro, A., Dasso, M., Lorca, T., Sicheri, F.*, and Pintard, L*. (2021). Bora phosphorylation substitutes in trans for T-loop phosphorylation in Aurora A to promote mitotic entry. Nat Commun 12, 1899. doi: 10.1038/s41467-021-21922-w
Velez-Aguilera, G., Nkombo Nkoula, S., Ossareh-Nazari, B., Link, J., Paouneskou, D., Van Hove, L., Joly, N., Tavernier, N., Verbavatz, J. M., Jantsch, V., and Pintard, L. (2020). PLK-1 promotes the merger of the parental genome into a single nucleus by triggering lamina disassembly. Elife 9, 9:e59510. doi: 10.7554/eLife.59510.
Joly, N.*, Beaumale, E., Van Hove, L., Martino, L., and Pintard, L*. (2020). Phosphorylation of the microtubule-severing AAA+ enzyme Katanin regulates C. elegans embryo development. J Cell Biol 219, (6):e201912037. doi: 10.1083/jcb.201912037.
Martino, L., Morchoisne-Bolhy, S., Cheerambathur, D. K., Van Hove, L., Dumont, J., Joly, N., Desai, A., Doye, V., and Pintard, L*. (2017). Channel Nucleoporins Recruit PLK-1 to Nuclear Pore Complexes to Direct Nuclear Envelope Breakdown in C. elegans. Dev Cell 43(2):157-171.e7. doi: 10.1016/j.devcel.2017.09.019.
* Corresponding authors
Publications
Strzelecki, P., Joly, N., Hébraud, P., Hoffmann, E., Cech, G. M., Kloska, A., Busi, F., & Grange, W. (2024). Enhanced Golden Gate Assembly: evaluating overhang strength for improved ligation efficiency. Nucleic Acids Research, gkae809. https://doi.org/10.1093/nar/gkae809
Beaumale, E., Van Hove, L., Pintard, L., & Joly, N. (2024). Microtubule-binding domains in Katanin p80 subunit are essential for severing activity in C. elegans. Journal of Cell Biology, 223(4), e202308023. https://doi.org/10.1083/jcb.202308023
Nkombo Nkoula, S., Velez-Aguilera, G., Ossareh-Nazari, B., Van Hove, L., Ayuso, C., Legros, V., Chevreux, G., Thomas, L., Seydoux, G., Askjaer, P., & Pintard, L. (2023). Mechanisms of nuclear pore complex disassembly by the mitotic Polo-like kinase 1 (PLK-1) in C. elegans embryos. Science Advances, 9(29), eadf7826. https://doi.org/10.1126/sciadv.adf7826
Kouranti, I., Abdel Khalek, W., Mazurkiewicz, S., Loisel-Ferreira, I., Gautreau, A. M., Pintard, L., Jeunemaitre, X., & Clauser, E. (2022). Cullin 3 Exon 9 Deletion in Familial Hyperkalemic Hypertension Impairs Cullin3-Ring-E3 Ligase (CRL3) Dynamic Regulation and Cycling. International Journal of Molecular Sciences, 23(9), 5151. https://doi.org/10.3390/ijms23095151
Velez-Aguilera, G., Ossareh-Nazari, B., Van Hove, L., Joly, N., & Pintard, L. (2022). Cortical microtubule pulling forces contribute to the union of the parental genomes in the Caenorhabditis elegans zygote. ELife, 11, e75382. https://doi.org/10.7554/eLife.75382
Liu, D., Marie, J.-C., Pelletier, A.-L., Song, Z., Ben-Khemis, M., Boudiaf, K., Pintard, C., Leger, T., Terrier, S., Chevreux, G., El-Benna, J., & Dang, P. M.-C. (2022). Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation. Cellular and Molecular Gastroenterology and Hepatology, 13(4), 1073–1093. https://doi.org/10.1016/j.jcmgh.2022.01.003
Knox, J., Joly, N., Linossi, E. M., Carmona-Negrón, J. A., Jura, N., Pintard, L., Zuercher, W., & Roy, P. J. (2021). A survey of the kinome pharmacopeia reveals multiple scaffolds and targets for the development of novel anthelmintics. Scientific Reports, 11(1), 9161. https://doi.org/10.1038/s41598-021-88150-6
Tavernier, N., Thomas, Y., Vigneron, S., Maisonneuve, P., Orlicky, S., Mader, P., Regmi, S. G., Van Hove, L., Levinson, N. M., Gasmi-Seabrook, G., Joly, N., Poteau, M., Velez-Aguilera, G., Gavet, O., Castro, A., Dasso, M., Lorca, T., Sicheri, F., & Pintard, L. (2021). Bora phosphorylation substitutes in trans for T-loop phosphorylation in Aurora A to promote mitotic entry. Nature Communications, 12(1), 1899. https://doi.org/10.1038/s41467-021-21922-w
Velez-Aguilera, G., Nkombo Nkoula, S., Ossareh-Nazari, B., Link, J., Paouneskou, D., Van Hove, L., Joly, N., Tavernier, N., Verbavatz, J.-M., Jantsch, V., & Pintard, L. (2020). PLK-1 promotes the merger of the parental genome into a single nucleus by triggering lamina disassembly. ELife, 9, e59510. https://doi.org/10.7554/eLife.59510
Joly, N., Beaumale, E., Van Hove, L., Martino, L., & Pintard, L. (2020). Phosphorylation of the microtubule-severing AAA+ enzyme Katanin regulates C. elegans embryo development. The Journal of Cell Biology, 219(6), e201912037. https://doi.org/10.1083/jcb.201912037
Gutnik, S., Thomas, Y., Guo, Y., Stoecklin, J., Neagu, A., Pintard, L., Merlet, J., & Ciosk, R. (2018). PRP-19, a conserved pre-mRNA processing factor and E3 ubiquitin ligase, inhibits the nuclear accumulation of GLP-1/Notch intracellular domain. Biology Open, 7(7), bio034066. https://doi.org/10.1242/bio.034066
Vigneron, S., Sundermann, L., Labbé, J.-C., Pintard, L., Radulescu, O., Castro, A., & Lorca, T. (2018). Cyclin A-cdk1-Dependent Phosphorylation of Bora Is the Triggering Factor Promoting Mitotic Entry. Developmental Cell, 45(5), 637-650.e7. https://doi.org/10.1016/j.devcel.2018.05.005
Martino, L., Morchoisne-Bolhy, S., Cheerambathur, D. K., Van Hove, L., Dumont, J., Joly, N., Desai, A., Doye, V., & Pintard, L. (2017). Channel Nucleoporins Recruit PLK-1 to Nuclear Pore Complexes to Direct Nuclear Envelope Breakdown in C. elegans. Developmental Cell, 43(2), 157-171.e7. https://doi.org/10.1016/j.devcel.2017.09.019
Dickinson, D. J., Schwager, F., Pintard, L., Gotta, M., & Goldstein, B. (2017). A Single-Cell Biochemistry Approach Reveals PAR Complex Dynamics during Cell Polarization. Developmental Cell, 42(4), 416-434.e11. https://doi.org/10.1016/j.devcel.2017.07.024
Richarme, G., Liu, C., Mihoub, M., Abdallah, J., Leger, T., Joly, N., Liebart, J.-C., Jurkunas, U. V., Nadal, M., Bouloc, P., Dairou, J., & Lamouri, A. (2017). Guanine glycation repair by DJ-1/Park7 and its bacterial homologs. Science (New York, N.Y.), 357(6347), 208–211. https://doi.org/10.1126/science.aag1095
Reviews
Book chapters
- Frank SICHERI (University of Toronto, Canada)
- Thierry LORCA (CRBM, Montpellier, France)
- Anna CASTRO (CRBM, Montpellier, France)
- Olivier GAVET (Institut Gustave Roussy, Paris Villejuif)
- Mary DASSO (NIH, Bethesda USA)
- Peter ASKJAER (CABD, Sevilla, Spain)
- Verena JANTSCH (Max Perutz Labs Vienna, Austria)
- Antoine JEGOU (Institut Jacques Monod, Paris, France)
- Guillaume ROMET-LEMONNE (Institut Jacques Monod, Paris, France)
- Denis CHRETIEN (IGDR, Rennes, France)
- Julien DUMONT (Institut Jacques Monod, Paris, France)
- Valérie DOYE (Institut Jacques Monod, Paris, France)
- Bruce BOWERMAN (IMB, University of Oregon, USA)
- Arshad DESAI (USCD, San Diego, USA)
- Monica GOTTA (University of Geneva, Switzerland)
- ANR AMBRE
- ANR REPLIGREAT
- ARC
- Idex « AAP Dynamique » Université de Paris
- Equipe Labellisée Ligue contre le Cancer
- Labex « WHO AM I »
25/10/2021 : Nicolas JOLY (Institut Jacques Monod), winner of the Maurice Nicloux Prize 2021!
Master and PhD students
Our laboratory offers a wide range of interesting and challenging research projects for motivated master or PhD candidates aimed at understanding the mechanisms of cell cycle control during animal development.
We employ a unique combination of genetics, biochemistry, cell biology, live imaging, quantitative proteomics, and functional genomics approaches.
PostDoc candidates
We are looking for highly motivated and team-oriented scientists with a strong background in biochemistry, genetics and cell biology.
Candidates are welcome to apply with CV, publication list, motivation letter, and names of 2 referees to:
Lionel PINTARD – Université Paris cité, CNRS, Institut Jacques Monod, 15 rue Hélène Brion – 75013 PARIS cedex 13 – France