RNF212B E3 ligase is essential for crossover designation and maturation during male and female meiosis in the mouse

Proceedings of the National Academy of Sciences, vol. 121, pp. e2320995121

Abstract

Meiosis, a reductional cell division, relies on precise initiation, maturation, and resolution of crossovers (COs) during prophase I to ensure the accurate segregation of homologous chromosomes during metaphase I. This process is regulated by the interplay of RING-E3 ligases such as RNF212 and HEI10 in mammals. In this study, we functionally characterized a recently identified RING-E3 ligase, RNF212B. RNF212B colocalizes and interacts with RNF212, forming foci along chromosomes from zygonema onward in a synapsis-dependent and DSB-independent manner. These consolidate into larger foci at maturing COs, colocalizing with HEI10, CNTD1, and MLH1 by late pachynema. Genetically, RNF212B foci formation depends on Rnf212 but not on Msh4 , Hei10, and Cntd1 , while the unloading of RNF212B at the end of pachynema is dependent on Hei10 and Cntd1 . Mice lacking RNF212B, or expressing an inactive RNF212B protein, exhibit modest synapsis defects, a reduction in the localization of pro-CO factors (MSH4, TEX11, RPA, MZIP2) and absence of late CO-intermediates (MLH1). This loss of most COs by diakinesis results in mostly univalent chromosomes. Double mutants for Rnf212b and Rnf212 exhibit an identical phenotype to that of Rnf212b single mutants, while double heterozygous demonstrate a dosage-dependent reduction in CO number, indicating a functional interplay between paralogs. SUMOylome analysis of testes from Rnf212b mutants and pull-down analysis of Sumo- and Ubiquitin-tagged HeLa cells, suggest that RNF212B is an E3-ligase with Ubiquitin activity, serving as a crucial factor for CO maturation. Thus, RNF212 and RNF212B play vital, yet overlapping roles, in ensuring CO homeostasis through their distinct E3 ligase activities.

Authors 17

1. Yazmine B. Condezo corresponding

   Universidad de Salamanca · Centro de Investigación del Cáncer · Instituto de Biología Molecular y Celular del Cáncer de Salamanca

   Affiliation as printed

   Molecular Mechanisms Program, Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (onsejo Superior de Investigaciones Científicas-Universidad de Salamanca), 37007 Salamanca, Spain

2. Raquel Sainz-Urruela corresponding

   Universidad de Salamanca · Centro de Investigación del Cáncer · Instituto de Biología Molecular y Celular del Cáncer de Salamanca

   Affiliation as printed

   Molecular Mechanisms Program, Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (onsejo Superior de Investigaciones Científicas-Universidad de Salamanca), 37007 Salamanca, Spain

3. Laura Gómez-H corresponding

   Universidad de Salamanca · Centro de Investigación del Cáncer · Max Planck Institute of Biochemistry · Instituto de Biología Molecular y Celular del Cáncer de Salamanca

   Affiliation as printed

   Department of Totipotency, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany

   Molecular Mechanisms Program, Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (onsejo Superior de Investigaciones Científicas-Universidad de Salamanca), 37007 Salamanca, Spain

4. Daniel Salas‐Lloret Aachen Department of Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Department of Cell and Chemical Biology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands

5. Natalia Felipe‐Medina

   Universidad de Salamanca · Centro de Investigación del Cáncer · Instituto de Biología Molecular y Celular del Cáncer de Salamanca

   Affiliation as printed

   Molecular Mechanisms Program, Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (onsejo Superior de Investigaciones Científicas-Universidad de Salamanca), 37007 Salamanca, Spain

6. Rachel A. Bradley

   Cornell University

   Affiliation as printed

   Department of Biomedical Sciences, Cornell University, Ithaca, NY 14853

7. Ian D. Wolff

   Cornell University

   Affiliation as printed

   Department of Biomedical Sciences, Cornell University, Ithaca, NY 14853

8. Stephanie Tanis

   Cornell University

   Affiliation as printed

   Department of Biomedical Sciences, Cornell University, Ithaca, NY 14853

9. José Luís Barbero

   Consejo Superior de Investigaciones Científicas · Centro de Investigaciones Biológicas Margarita Salas

   Affiliation as printed

   Centro de Investigaciones Biológicas Margarita Salas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, Spain

10. Manuel Adolfo Sánchez-Martin

    Universidad de Salamanca

    Affiliation as printed

    Departamento de Medicina, Universidad de Salamanca, 37007 Salamanca, Spain

11. Dirk G. de Rooij

    Utrecht University

    Affiliation as printed

    Reproductive Biology Group, Division of Developmental Biology, Department of Biology, Faculty of Science, Utrecht University, Utrecht 3584CM, The Netherlands

12. Ivo Alexander Hendriks

    University of Copenhagen · Novo Nordisk Foundation

    Affiliation as printed

    Proteomics program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark

13. Michael Lund Nielsen

    University of Copenhagen · Novo Nordisk Foundation

    Affiliation as printed

    Proteomics program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark

14. Román González‐Prieto Aachen Department of Cell and Chemical Biology

    Consejo Superior de Investigaciones Científicas · Leiden University Medical Center · Centro Andaluz de Biología Molecular y Medicina Regenerativa · Universidad de Sevilla · Universidad Pablo de Olavide

    Affiliation as printed

    Andalusian Center for Molecular Biology and Regenerative MedicineCentro Andaluz de Biología Molecular y Medicina Regenerativa, Universidad de Sevilla-Consejo Superior de Investigaciones Científicas-Universidad-Pablo de Olavide, 41092 Sevilla, Spain

    Departamento de Biología Celular, Facultad de Biología, Universidad de Sevilla, 41012 Sevilla, Spain

    Department of Cell and Chemical Biology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands

15. Paula Elaine Cohen

    Cornell University

    Affiliation as printed

    Department of Biomedical Sciences, Cornell University, Ithaca, NY 14853

16. Alberto M. Pendás corresponding

    Universidad de Salamanca · Centro de Investigación del Cáncer · Instituto de Biología Molecular y Celular del Cáncer de Salamanca

    Affiliation as printed

    Molecular Mechanisms Program, Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (onsejo Superior de Investigaciones Científicas-Universidad de Salamanca), 37007 Salamanca, Spain

17. Elena Llano corresponding

    Universidad de Salamanca · Centro de Investigación del Cáncer · Instituto de Biología Molecular y Celular del Cáncer de Salamanca

    Affiliation as printed

    Departamento de Fisiología, Universidad de Salamanca, 37007 Salamanca, Spain

    Molecular Mechanisms Program, Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (onsejo Superior de Investigaciones Científicas-Universidad de Salamanca), 37007 Salamanca, Spain