Abstract
The hexameric AAA ATPase p97 is involved in several human proteinopathies and mediates ubiquitin-dependent protein degradation among other essential cellular processes. Via its N-terminal domain (N domain), p97 interacts with multiple regulatory cofactors including the UFD1/NPL4 heterodimer and members of the "ubiquitin regulatory X" (UBX) domain protein family; however, the principles governing cofactor selectivity remain to be deciphered. Our crystal structure of the FAS-associated factor 1 (FAF1)UBX domain in complex with the p97N domain reveals that the signature Phe-Pro-Arg motif known to be crucial for interactions of UBX domains with p97 adopts a cis-proline configuration, in contrast to a cis-trans mixture we derive for the isolated FAF1UBX domain. Biochemical studies confirm that binding critically depends on a proline at this position. Furthermore, we observe that the UBX proteins FAF1 and UBXD7 only bind to p97-UFD1/NPL4, but not free p97, thus demonstrating for the first time a hierarchy in p97-cofactor interactions.
Copyright © 2011 Elsevier Ltd. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / chemistry*
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Adaptor Proteins, Vesicular Transport
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Adenosine Triphosphatases / chemistry*
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Amino Acid Motifs
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Amino Acid Sequence
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Apoptosis Regulatory Proteins
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Calorimetry
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Carrier Proteins / chemistry
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Chromatography, Gel
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Coenzymes / chemistry*
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Conserved Sequence
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Crystallography, X-Ray
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Humans
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Intracellular Signaling Peptides and Proteins
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Molecular Sequence Data
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Nuclear Proteins / chemistry*
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Protein Binding
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Protein Interaction Domains and Motifs
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Protein Structure, Quaternary
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Protein Structure, Tertiary
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Proteins / chemistry
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Surface Properties
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Titrimetry
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Ubiquitin / chemistry
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Ubiquitinated Proteins / chemistry
Substances
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Adaptor Proteins, Signal Transducing
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Adaptor Proteins, Vesicular Transport
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Apoptosis Regulatory Proteins
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Carrier Proteins
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Coenzymes
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FAF1 protein, human
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Intracellular Signaling Peptides and Proteins
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NPLOC4 protein, human
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Nuclear Proteins
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Proteins
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UBXN7 protein, human
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UFD1 protein, human
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Ubiquitin
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Ubiquitinated Proteins
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Adenosine Triphosphatases
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p97 ATPase
Associated data
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PDB/3QQ7
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PDB/3QQ8
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PDB/3R3M