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Proteasome recruitment and activation of the Uch37 deubiquitinating enzyme by Adrm1

Nature Cell Biology volume 8, pages 994–1002 (2006)Cite this article

Abstract

Uch37 is one of the three principal deubiquitinating enzymes (DUBs), and the only ubiquitin carboxy-terminal hydrolase (UCH)-family protease, that is associated with mammalian proteasomes. We show that Uch37 is responsible for the ubiquitin isopeptidase activity in the PA700 (19S) proteasome regulatory complex1. PA700 isopeptidase disassembles Lys 48-linked polyubiquitin specifically from the distal end of the chain, a property that may be used to clear poorly ubiquitinated or unproductively bound substrates from the proteasome. To better understand Uch37 function and the mechanism responsible for its specificity, we investigated how Uch37 is recruited to proteasomes. Uch37 binds through Adrm1, a previously unrecognized orthologue of Saccharomyces cerevisiae Rpn13p, which in turn is bound to the S1 (also known as Rpn2) subunit of the 19S complex. Adrm1 (human Rpn13, hRpn13) binds the carboxy-terminal tail of Uch37, a region that is distinct from the UCH catalytic domain, which we show inhibits Uch37 activity. Following binding, Adrm1 relieves Uch37 autoinhibition, accelerating the hydrolysis of ubiquitin-7-amido-4-methylcoumarin (ubiquitin−AMC). However, neither Uch37 alone nor the Uch37–Adrm1 or Uch37–Adrm1–S1 complexes can hydrolyse di-ubiquitin efficiently; rather, incorporation into the 19S complex is required to enable processing of polyubiquitin chains.

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Figure 1: Human Uch37 is associated with the 26S proteasome in vivo and disassembles di-ubiquitin.
Figure 2: Adrm1 interacts with Uch37 in vivo and in vitro.
Figure 3: Adrm1 is a human homologue of yeast Rpn13p and a subunit of the proteasome.
Figure 4: Adrm1 recruits Uch37 to the proteasome.
Figure 5: Deubiquitination by Uch37 is activated by Adrm1 and in the 19S regulatory complex.

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Acknowledgements

We thank C. Gorbea and M. Rechsteiner for plasmids containing 19S complex cDNAs, A. Borodovsky and H. Ploegh for UbVS, B. Plapp for advice on fitting kinetic data and C. Slaughter for mass spectrometry of Uch37 peptides. This work was supported, in part, by the National Institutes of Health grants R01 GM37666 (to R.E.C.) and R37 GM41628 (to R.C.C.). T.Y. is a fellow of the Leukemia and Lymphoma Society.

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Author notes

  1. Wei Xu

    Present address: McArdle Laboratory for Cancer Research, Madison, WI 53706, USA

  2. Robert E. Cohen

    Present address: Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA

Authors and Affiliations

  1. Stowers Institute for Medical Research, Kansas City, 64110, MO, USA

    Tingting Yao, Laurence Florens, Selene K. Swanson, Michael P. Washburn, Ronald C. Conaway & Joan Weliky Conaway

  2. Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, 52242, IA, USA

    Tingting Yao, Ling Song, Wei Xu & Robert E. Cohen

  3. Department of Physiology, University of Texas Southwestern Medical Center, Dallas, 75390, TX, USA

    George N. DeMartino

  4. Department of Biochemistry and Molecular Biology, Kansas University Medical Center, Kansas City, 66160, KS, USA

    Ronald C. Conaway & Joan Weliky Conaway

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Supplementary Figures S1, S2, S3 and Supplementary Table S1 (PDF 1563 kb)

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Yao, T., Song, L., Xu, W. et al. Proteasome recruitment and activation of the Uch37 deubiquitinating enzyme by Adrm1. Nat Cell Biol 8, 994–1002 (2006). https://doi.org/10.1038/ncb1460

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