Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15

Teyra, J.; Singer, A.; Schmitges, F.; Jaynes, P.; Kit Leng Lui, S.; Polyak, M.; Fodil, N.; Krieger, J.; Tong, J.; Schwerdtfeger, C.; Brasher, B.; Ceccarelli, DFJ; Moffat, J.; Sicheri, F.; Moran, M.; Gros, P.; Eichhorn, Pieter; Lenter, M.; Boehmelt, G.; Sidhu, S.

doi:10.1016/j.str.2019.01.002

Access Status

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Authors

Teyra, J.

Singer, A.

Schmitges, F.

Jaynes, P.

Kit Leng Lui, S.

Polyak, M.

Fodil, N.

Krieger, J.

Tong, J.

Schwerdtfeger, C.

Brasher, B.

Ceccarelli, DFJ

Moffat, J.

Sicheri, F.

Moran, M.

Gros, P.

Eichhorn, Pieter

Lenter, M.

Boehmelt, G.

Sidhu, S.

Date

2019

Type

Journal Article

Metadata

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Citation

Teyra, J. and Singer, A.U. and Schmitges, F.W. and Jaynes, P. and Kit Leng Lui, S. and Polyak, M.J. and Fodil, N. et al. 2019. Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15. Structure. 27 (4): pp. 590-605.e5.

Source Title

Structure

DOI

10.1016/j.str.2019.01.002

ISSN

0969-2126

Faculty

Faculty of Health Sciences

School

School of Pharmacy and Biomedical Sciences

URI

http://hdl.handle.net/20.500.11937/75729

Collection

Curtin Research Publications

Abstract

The multi-domain deubiquitinase USP15 regulates diverse eukaryotic processes and has been implicated in numerous diseases. We developed ubiquitin variants (UbVs) that targeted either the catalytic domain or each of three adaptor domains in USP15, including the N-terminal DUSP domain. We also designed a linear dimer (diUbV), which targeted the DUSP and catalytic domains, and exhibited enhanced specificity and more potent inhibition of catalytic activity than either UbV alone. In cells, the UbVs inhibited the deubiquitination of two USP15 substrates, SMURF2 and TRIM25, and the diUbV inhibited the effects of USP15 on the transforming growth factor β pathway. Structural analyses revealed that three distinct UbVs bound to the catalytic domain and locked the active site in a closed, inactive conformation, and one UbV formed an unusual strand-swapped dimer and bound two DUSP domains simultaneously. These inhibitors will enable the study of USP15 function in oncology, neurology, immunology, and inflammation.