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Dissecting the Multiple Functions of the Polo-Like Kinase 1 in the C. elegans Zygote

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Cell Cycle Control

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2740))

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Abstract

Plk1 (polo-like kinase 1) is an evolutionarily conserved serine/threonine kinase instrumental for mitotic entry and progression. Beyond these canonical functions, Plk1 also regulates cell polarization and cell fate during asymmetric cell divisions in C. elegans and D. melanogaster. Plk1 contains a specialized phosphoserine–threonine binding domain, the polo-box domain (PBD), which localizes and concentrates the kinase at its various sites of action within the cell in space and time. Here we present protocols to express and purify the C. elegans Plk1 kinase along with biochemical and phosphoproteomic approaches to interrogate the PBD interactome and to dissect Plk1 substrate interactions. These protocols are most suitable for the identification of Plk1 targets in C. elegans embryos but can be easily adapted to identify and study Plk1 substrates from any source.”

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Acknowledgments

We thank P. Moussounda for media preparation and Véronique Legros and Guillaume Chevreux, from the Proteomic platform, for phosphopeptide purification and tandem mass spectrometry analysis. We thank R. Karess for critical reading of the manuscript. We thank all members of the team for stimulating discussions. Work in LP laboratory is supported by grants from Agence Nationale pour la Recherche (ANR), France, ANR-22-CE13-0022 (LP), La Ligue Nationale Contre le Cancer, Equipe Labellisée, France, and Idex Université Paris Cité, ANR-18-IDEX-0001. Griselda Velez-Aguilera is supported by a postdoctoral fellowship from “Secretaría de Educación, Ciencia, Tecnología e Innovación de la Ciudad de México,” CMSECTEI/201/2022.

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Author notes

  1. Griselda Velez-Aguilera and Batool Ossareh-Nazari, have equally contributed to this chapter.

Authors and Affiliations

  1. Université Paris Cité, CNRS, Institut Jacques Monod, Paris, France

    Griselda Velez-Aguilera, Batool Ossareh-Nazari & Lionel Pintard

  2. Programme Equipe Labellisée Ligue Contre le Cancer, Paris, France

    Griselda Velez-Aguilera, Batool Ossareh-Nazari & Lionel Pintard

Authors
  1. Griselda Velez-Aguilera
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  2. Batool Ossareh-Nazari
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  3. Lionel Pintard
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Corresponding author

Correspondence to Lionel Pintard .

Editor information

Editors and Affiliations

  1. Université de Montpellier, Centre de Recherche en Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, Montpellier Cedex 5, France

    Anna Castro

  2. Université de Montpellier, Centre de Recherche en Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, Montpellier Cedex 5, France

    Benjamin Lacroix

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Velez-Aguilera, G., Ossareh-Nazari, B., Pintard, L. (2024). Dissecting the Multiple Functions of the Polo-Like Kinase 1 in the C. elegans Zygote. In: Castro, A., Lacroix, B. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 2740. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3557-5_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3557-5_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3556-8

  • Online ISBN: 978-1-0716-3557-5

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