Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15

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 designe...

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Main 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.
Format: Journal Article
Language:English
Published: CELL PRESS 2019
Subjects:
Online Access:http://hdl.handle.net/20.500.11937/75729
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author 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.
author_facet 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.
author_sort Teyra, J.
building Curtin Institutional Repository
collection Online Access
description 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.
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spelling curtin-20.500.11937-757292019-06-11T04:09:46Z 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. Science & Technology Life Sciences & Biomedicine Biochemistry & Molecular Biology Biophysics Cell Biology CELL-CYCLE DEUBIQUITYLATING ENZYMES DEUBIQUITINATING ENZYMES E3 LIGASES PHOSPHORYLATION SPECIFICITY RECOGNITION CENTROSOME MODULATION DISCOVERY 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. 2019 Journal Article http://hdl.handle.net/20.500.11937/75729 10.1016/j.str.2019.01.002 English CELL PRESS restricted
spellingShingle Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Biophysics
Cell Biology
CELL-CYCLE
DEUBIQUITYLATING ENZYMES
DEUBIQUITINATING ENZYMES
E3 LIGASES
PHOSPHORYLATION
SPECIFICITY
RECOGNITION
CENTROSOME
MODULATION
DISCOVERY
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.
Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
title Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
title_full Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
title_fullStr Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
title_full_unstemmed Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
title_short Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
title_sort structural and functional characterization of ubiquitin variant inhibitors of usp15
topic Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Biophysics
Cell Biology
CELL-CYCLE
DEUBIQUITYLATING ENZYMES
DEUBIQUITINATING ENZYMES
E3 LIGASES
PHOSPHORYLATION
SPECIFICITY
RECOGNITION
CENTROSOME
MODULATION
DISCOVERY
url http://hdl.handle.net/20.500.11937/75729