Quantitative interaction mapping reveals an extended UBX domain in ASPL that disrupts functional p97 hexamers.

Interaction mapping is a powerful strategy to elucidate the biological function of protein assemblies and their regulators. Here, we report the generation of a quantitative interaction network, directly linking 14 human proteins to the AAA+ ATPase p97, an essential hexameric protein with multiple cellular functions. We show that the high-affinity ...
interacting protein ASPL efficiently promotes p97 hexamer disassembly, resulting in the formation of stable p97:ASPL heterotetramers. High-resolution structural and biochemical studies indicate that an extended UBX domain (eUBX) in ASPL is critical for p97 hexamer disassembly and facilitates the assembly of p97:ASPL heterotetramers. This spontaneous process is accompanied by a reorientation of the D2 ATPase domain in p97 and a loss of its activity. Finally, we demonstrate that overproduction of ASPL disrupts p97 hexamer function in ERAD and that engineered eUBX polypeptides can induce cell death, providing a rationale for developing anti-cancer polypeptide inhibitors that may target p97 activity.
Mesh Terms:
Brain, Cell Proliferation, Crystallography, X-Ray, Endoplasmic Reticulum-Associated Degradation, HEK293 Cells, Humans, Mutation, Oncogene Proteins, Fusion, Peptides, Protein Binding, Protein Domains, Protein Engineering, Protein Interaction Maps, Protein Multimerization, Protein Structure, Quaternary, Recombinant Proteins, Valosin Containing Protein
Nat Commun
Date: Dec. 20, 2015
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