Abstract

Thiolutin is a disulfide-containing antibiotic and anti-angiogenic compound produced by Streptomyces. Its biological targets are not known. We show that reduced thiolutin is a zinc chelator that inhibits the JAB1/MPN/Mov34 (JAMM) domain–containing metalloprotease Rpn11, a deubiquitinating enzyme of the 19S proteasome. Thiolutin also inhibits the JAMM metalloproteases Csn5, the deneddylase of the COP9 signalosome; AMSH, which regulates ubiquitin-dependent sorting of cell-surface receptors; and BRCC36, a K63-specific deubiquitinase of the BRCC36-containing isopeptidase complex and the BRCA1–BRCA2-containing complex. We provide evidence that other dithiolopyrrolones also function as inhibitors of JAMM metalloproteases.

  • Compound

    PYR2604

  • Compound

    Holomycin

  • Compound

    Methacryloyl holothin

  • Compound

    Crotonyl holothin

  • Compound

    3,3-Dimethylacryloyl holothin

  • Compound

    Tigloyl holothin

  • Compound

    Pentanoyl holothin

  • Compound

    5-Hexynoyl holothin

  • Compound

    Hexanoyl holothin

  • Compound

    Octanoyl holothin

  • Compound

    6-Azidohexanoyl holothin

  • Compound

    Benzoyl holothin

  • Compound

    Pivaloyl holothin

  • Compound

    Pseudomonyl C holothinamide

  • Compound

    Thiomarinol A

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Acknowledgements

We thank T.J. Simpson (University of Bristol) for thiomarinol, S. Cohen (University of California, San Diego) for Zn2+-cyclen, E. Zeqiraj (University of Leeds) for BRISC complex, D. Marzoll (Heidelberg University Biochemistry Center) for purified recombinant Flag-FRQ, A. Martin (University of California, Berkeley) for the pETDuet-1 vector expressing the Rpn11–Rpn8 dimer and M. Knop (ZMBH Heidelberg) for yeast strains. The work was supported by Sonderforschungsbereich 1036 grants to M.B. and F.M., Transregio 186 grant to M.B., Deutsche Forschungsgemeinschaft grant DI1874/1 to A.D. and US National Institutes of Health grant CA164803 to R.J.D. F.M. and M.B. are investigators of Cellnetworks. R.J.D. is an investigator of and was supported by the Howard Hughes Medical Institute.

Author information

Author notes

    • Linda Lauinger
    • , Nati Ha
    • , Philipp E Merkl
    •  & Tobias Schafmeier

    Present addresses: University of California, Irvine, School of Medicine, Department of Biological Chemistry, Irvine, California, USA (L.L.); German Cancer Research Center (DKFZ), Applied Bioinformatics, Heidelberg, Germany (N.H.); Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA (P.E.M.); Institute for Diabetes and Cancer (IDC) Helmholtz Center Munich, Neuherberg, Germany (T.S.).

Affiliations

  1. Heidelberg University Biochemistry Center, Heidelberg, Germany.

    • Linda Lauinger
    • , Anton Shostak
    • , Ibrahim Avi Cemel
    • , Nati Ha
    • , Tobias Schafmeier
    • , Michael Brunner
    •  & Axel Diernfellner
  2. Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA.

    • Jing Li
    • , Yaru Zhang
    •  & Raymond J Deshaies
  3. Lehrstuhl Biochemie III, Biochemie Zentrum Regensburg, Universität Regensburg, Regensburg, Germany.

    • Philipp E Merkl
    • , Simon Obermeyer
    •  & Herbert Tschochner
  4. Zentrum Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Alliance, Heidelberg University, Heidelberg, Germany.

    • Nicolas Stankovic-Valentin
    •  & Frauke Melchior
  5. Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

    • Walter J Wever
    •  & Albert A Bowers
  6. Department of Chemistry and Biochemistry, BioFrontiers Institute, University of Colorado Boulder, Boulder, Colorado, USA.

    • Kyle P Carter
    •  & Amy E Palmer
  7. Howard Hughes Medical Institute, California Institute of Technology, Pasadena, California, USA.

    • Raymond J Deshaies

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Contributions

A.D. and M.B. designed the research and wrote the manuscript; J.L., Y.Z. and R.J.D. designed, performed and interpreted the Rpn11, Csn5, AMSH and BRCC36 experiments. W.J.W. and A.A.B. synthesized the holothin derivatives. K.P.C. and A.E.P. designed and performed the measurements of intracellular zinc. N.S.-V. and F.M. designed and performed the USP5 and USP25 experiments; P.E.M., S.O. and H.T. designed and performed the Pol I–III in vitro transcription assays. L.L., I.A.C., A.D. and T.S. performed the Neurospora experiments. L.L. and N.H. performed the RNA-seq analysis. L.L. performed the yeast experiments. A.S. and L.L. performed the HeLa experiments. R.J.D. implicated THL as an Rpn11 inhibitor and edited the manuscript.

Competing interests

R.J.D. is a founder, shareholder, member of the scientific advisory board and consultant of Cleave Biosciences, which is developing drugs that target protein homeostasis in cancer.

Corresponding authors

Correspondence to Michael Brunner or Axel Diernfellner.

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    Supplementary Results, Supplementary Table 1 and Supplementary Figures 1–10

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    Synthesis and Characterization of Chemical Compounds

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DOI

https://doi.org/10.1038/nchembio.2370