Arginine (R)-based ER localization signals are sorting motifs that confer transient ER localization to unassembled subunits of multimeric membrane proteins. The COPI vesicle coat binds R-based signals but the molecular details remain unknown. Here, we use reporter membrane proteins based on the proteolipid Pmp2 fused to GFP and allele swapping of COPI subunits to map the recognition site for R-based signals. We show that two highly conserved stretches—in the β- and δ-COPI subunits—are required to maintain Pmp2GFP reporters exposing R-based signals in the ER. Combining a deletion of 21 residues in δ-COP together with the mutation of three residues in β-COP gave rise to a COPI coat that had lost its ability to recognize R-based signals, whilst the recognition of C-terminal di-lysine signals remained unimpaired. A homology model of the COPI trunk domain illustrates the recognition of R-based signals by COPI.
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22 October 2007
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October 22 2007
Novel cargo-binding site in the β and δ subunits of coatomer
Kai Michelsen,
Kai Michelsen
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Volker Schmid,
Volker Schmid
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Jutta Metz,
Jutta Metz
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Katja Heusser,
Katja Heusser
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Urban Liebel,
Urban Liebel
2European Molecular Biology Laboratory (EMBL), Cell Biology Cell Biophysics Unit, D-69117 Heidelberg, Germany
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Torsten Schwede,
Torsten Schwede
3Swiss Institute of Bioinformatics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland
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Anne Spang,
Anne Spang
4Growth & Development, Biozentrum, University of Basel, CH-4056 Basel, Switzerland
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Blanche Schwappach
Blanche Schwappach
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Kai Michelsen
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
Volker Schmid
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
Jutta Metz
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
Katja Heusser
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
Urban Liebel
2European Molecular Biology Laboratory (EMBL), Cell Biology Cell Biophysics Unit, D-69117 Heidelberg, Germany
Torsten Schwede
3Swiss Institute of Bioinformatics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland
Anne Spang
4Growth & Development, Biozentrum, University of Basel, CH-4056 Basel, Switzerland
Blanche Schwappach
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
Correspondence to Blanche Schwappach: [email protected]
U. Liebel's present address is Institute of Toxicology and Genetics, Forschungszentrum Karlsruhe, D-76021 Karlsruhe, Germany.
Abbreviations used in this paper: AD, activation domain; BD, binding domain.
Received:
April 24 2007
Accepted:
September 22 2007
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2007
J Cell Biol (2007) 179 (2): 209–217.
Article history
Received:
April 24 2007
Accepted:
September 22 2007
Citation
Kai Michelsen, Volker Schmid, Jutta Metz, Katja Heusser, Urban Liebel, Torsten Schwede, Anne Spang, Blanche Schwappach; Novel cargo-binding site in the β and δ subunits of coatomer . J Cell Biol 22 October 2007; 179 (2): 209–217. doi: https://doi.org/10.1083/jcb.200704142
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