Letter | Published:

Role of Cdc48/p97 as a SUMO-targeted segregase curbing Rad51–Rad52 interaction

Nature Cell Biology volume 15, pages 526532 (2013) | Download Citation

Abstract

Cdc48 (also known as p97), a conserved chaperone-like ATPase, plays a strategic role in the ubiquitin system1,2,3. Empowered by ATP-driven conformational changes 4, Cdc48 acts as a segregase by dislodging ubiquitylated proteins from their environment1,2,5. Ufd1, a known co-factor of Cdc48, also binds SUMO (ref. 6), but whether SUMOylated proteins are subject to the segregase activity of Cdc48 as well and what these substrates are remains unknown. Here we show that Cdc48 with its co-factor Ufd1 is SUMO-targeted to proteins involved in DNA double-strand break repair. Cdc48 associates with SUMOylated Rad52, a factor that assembles the Rad51 recombinase on chromatin. By acting on the Rad52–Rad51 complex, Cdc48 curbs their physical interaction and displaces the proteins from DNA. Genetically interfering with SUMO-targeting or segregase activity leads to an increase in spontaneous recombination rates, accompanied by aberrant in vivo Rad51 foci formation in yeast and mammalian cells. Our data thus suggest that SUMO-targeted Cdc48 restricts the recombinase Rad51 by counterbalancing the activity of Rad52. We propose that Cdc48, through its ability to associate with co-factors that have affinities for ubiquitin and SUMO, connects the two modification pathways for protein degradation or other regulatory purposes.

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Acknowledgements

We thank A. Strasser and K. Strasser for technical assistance and I. Psakhye for discussion; J. Gerlach for early contributions; and R. Kanaar (Erasmus MC, NL), S. C. Kowalczykowski (UC Davis, USA) and J. Lukas (CPR, DK) for materials. S.J. is supported by the Max Planck Society and the Louis-Jeantet Foundation; S.J. and S.B. by the RUBICON EU Network of Excellence; M.K. by Boehringer Ingelheim Fonds; S.J. and H.L. by Deutsche Forschungsgemeinschaft SFB646 and Centre for Integrated Protein Science Munich; L.S. and L.H. by Deutsche Forschungsgemeinschaft SFB TR5 and the Bioimaging Network Munich.

Author information

Author notes

    • Steven Bergink
    •  & Lothar Schermelleh

    Present addresses: Department of Cell Biology, University of Groningen, University Medical Center, Groningen 9713AV, The Netherlands (S.B.); Department of Biochemistry, University of Oxford, Oxford OX1 3Q4, UK (L.S.)

Affiliations

  1. Department of Molecular Cell Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany

    • Steven Bergink
    • , Tim Ammon
    • , Maximilian Kern
    •  & Stefan Jentsch
  2. Department of Biology and Center for Integrated Protein Science, Ludwig Maximilians University Munich, 82152 Martinsried, Germany

    • Lothar Schermelleh
    •  & Heinrich Leonhardt

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Contributions

S.B. and S.J. designed the experiments and obtained and analysed the data, T.A. contributed to the mammalian and M.K. the yeast part, L.S. and H.L. planned and conducted the super-resolution microscopy, S.J. and S.B. wrote the paper, and all authors contributed to the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Stefan Jentsch.

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    Three-dimensional reconstruction of abnormal filamentous Rad51 structures.

    Reconstruction of abnormal filamentous Rad51 structures accumulating in U2OS cells 12 h after induction of DNA damage after p97 RNAi treatment. Super-resolution fluorescence microscopy was done with the samples shown in Fig. 5b. The scale represents 1 μm.

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DOI

https://doi.org/10.1038/ncb2729

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