Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation

Jonas Boehringer; Christiane Riedinger; Konstantinos Paraskevopoulos; Eachan O D Johnson; Edward D Lowe; Christina Khoudian; Dominique Smith; Martin E M Noble; Colin Gordon; Jane A Endicott

doi:10.1042/BJ20120542

Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation

Jonas Boehringer, Christiane Riedinger, Konstantinos Paraskevopoulos, Eachan O D Johnson, Edward D Lowe, Christina Khoudian, Dominique Smith, Martin E M Noble, Colin Gordon, Jane A Endicott

MRC Human Genetics Unit

Research output: Contribution to journal › Article › peer-review

Abstract

The ubiquitin-proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein-protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.

Original language	English
Pages (from-to)	55-65
Number of pages	11
Journal	Biochemical Journal
Volume	448
Issue number	1
DOIs	https://doi.org/10.1042/BJ20120542
Publication status	Published - 15 Nov 2012

Keywords

Animals
Arabidopsis Proteins
Carrier Proteins
Circular Dichroism
Crystallography, X-Ray
Drosophila Proteins
Microtubule-Associated Proteins
Models, Molecular
Mutagenesis, Site-Directed
Peptide Fragments
Proteasome Endopeptidase Complex
Protein Binding
Protein Conformation
Protein Interaction Mapping
Protein Structure, Tertiary
Recombinant Proteins
Schizosaccharomyces
Schizosaccharomyces pombe Proteins
Structure-Activity Relationship
Ubiquitin
Winged-Helix Transcription Factors

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10.1042/BJ20120542

Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation
© 2012 The Author(s) The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited.
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Boehringer, Jonas ; Riedinger, Christiane ; Paraskevopoulos, Konstantinos ; Johnson, Eachan O D ; Lowe, Edward D ; Khoudian, Christina ; Smith, Dominique ; Noble, Martin E M ; Gordon, Colin ; Endicott, Jane A. / Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation. In: Biochemical Journal. 2012 ; Vol. 448, No. 1. pp. 55-65.

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abstract = "The ubiquitin-proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein-protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.",

keywords = "Animals, Arabidopsis Proteins, Carrier Proteins, Circular Dichroism, Crystallography, X-Ray, Drosophila Proteins, Microtubule-Associated Proteins, Models, Molecular, Mutagenesis, Site-Directed, Peptide Fragments, Proteasome Endopeptidase Complex, Protein Binding, Protein Conformation, Protein Interaction Mapping, Protein Structure, Tertiary, Recombinant Proteins, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Structure-Activity Relationship, Ubiquitin, Winged-Helix Transcription Factors",

author = "Jonas Boehringer and Christiane Riedinger and Konstantinos Paraskevopoulos and Johnson, {Eachan O D} and Lowe, {Edward D} and Christina Khoudian and Dominique Smith and Noble, {Martin E M} and Colin Gordon and Endicott, {Jane A}",

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Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation. / Boehringer, Jonas; Riedinger, Christiane; Paraskevopoulos, Konstantinos; Johnson, Eachan O D; Lowe, Edward D; Khoudian, Christina; Smith, Dominique; Noble, Martin E M; Gordon, Colin; Endicott, Jane A.

In: Biochemical Journal, Vol. 448, No. 1, 15.11.2012, p. 55-65.

Research output: Contribution to journal › Article › peer-review

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AU - Boehringer, Jonas

AU - Riedinger, Christiane

AU - Paraskevopoulos, Konstantinos

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AU - Lowe, Edward D

AU - Khoudian, Christina

AU - Smith, Dominique

AU - Noble, Martin E M

AU - Gordon, Colin

AU - Endicott, Jane A

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AB - The ubiquitin-proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein-protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.

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KW - Recombinant Proteins

KW - Schizosaccharomyces

KW - Schizosaccharomyces pombe Proteins

KW - Structure-Activity Relationship

KW - Ubiquitin

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JO - Biochemical Journal

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SN - 0264-6021

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Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation

Abstract

Keywords

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