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In Vitro Characterization of Sumoylation of HR Proteins

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Homologous Recombination

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2153))

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Abstract

In vitro analysis of posttranslational modifications such as sumoylation provides a great tool to not only identify the target proteins but also to characterize the specific effects of this modification on the protein features and uncover possible regulatory mechanism. In this chapter, we will describe the purification of yeast SUMO machinery proteins and their use to identify SUMO modification of target proteins in vitro. Furthermore, we will show several examples characterizing the effect of sumoylation on the biochemical activities of various proteins involved in homologous recombination (HR) that helped to better understand the regulatory role of this modification.

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Acknowledgments

We would like to thank Dr. Erica Johnson (Thomas Jefferson University) and Dr. Yoshiko Kikuchi (University of Tokyo) for providing expression plasmids. This work was supported by the Czech Science Foundation (GACR 17-17720s); the National Program of Sustainability II (MEYS CR, project no. LQ1605); the Wellcome trust Collaborative grant (206292/E/17/Z); and European Structural and Investment Funds, Operational Programme Research, Development and Education—“Preclinical Progression of New Organic Compounds with Targeted Biological Activity” (Preclinprogress)—CZ.02.1.01/0.0/0.0/16_025/0007381). We are grateful to Peter Kolesar for the critical reading of this manuscript.

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Authors and Affiliations

  1. Department of Biology, Masaryk University, Brno, Czech Republic

    Veronika Altmannova & Lumir Krejci

  2. International Clinical Research Center, St. Anne’s University Hospital, Brno, Czech Republic

    Veronika Altmannova & Lumir Krejci

  3. National Center for Biomolecular Research, Masaryk University, Brno, Czech Republic

    Lumir Krejci

Authors
  1. Veronika Altmannova
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  2. Lumir Krejci
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Correspondence to Veronika Altmannova or Lumir Krejci .

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Editors and Affiliations

  1. Centro Andaluz de Biologı´a Molecular, y Medicina Regenerativa, CABIMER, Universidad de Sevilla, Sevilla, Spain

    Andrés Aguilera

  2. Institut Curie, PSL Research University, CNRS, UMR3348, F-91405, Orsay, France

    Aura Carreira

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Altmannova, V., Krejci, L. (2021). In Vitro Characterization of Sumoylation of HR Proteins. In: Aguilera, A., Carreira, A. (eds) Homologous Recombination. Methods in Molecular Biology, vol 2153. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0644-5_33

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  • DOI: https://doi.org/10.1007/978-1-0716-0644-5_33

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0643-8

  • Online ISBN: 978-1-0716-0644-5

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