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Published December 3, 2019 | Accepted Version + Supplemental Material
Journal Article Open

Multisystem Proteinopathy Mutations in VCP/p97 Increase NPLOC4·UFD1L Binding and Substrate Processing


Valosin-containing protein (VCP)/p97 is an essential ATP-dependent protein unfoldase. Dominant mutations in p97 cause multisystem proteinopathy (MSP), a disease affecting the brain, muscle, and bone. Despite the identification of numerous pathways that are perturbed in MSP, the molecular-level defects of these p97 mutants are not completely understood. Here, we use biochemistry and cryoelectron microscopy to explore the effects of MSP mutations on the unfoldase activity of p97 in complex with its substrate adaptor NPLOC4⋅UFD1L (UN). We show that all seven analyzed MSP mutants unfold substrates faster. Mutant homo- and heterohexamers exhibit tighter UN binding and faster substrate processing. Our structural studies suggest that the increased UN affinity originates from a decoupling of p97's nucleotide state and the positioning of its N-terminal domains. Together, our data support a gain-of-function model for p97-UN-dependent processes in MSP and underscore the importance of N-terminal domain movements for adaptor recruitment and substrate processing by p97.

Additional Information

© 2019 Elsevier Ltd. Received 3 July 2019, Revised 19 August 2019, Accepted 20 September 2019, Available online 14 October 2019. We thank Rebecca Voorhees for advice on mammalian protein purification; Xiawei Huang for help with cloning; and the Deshaies and Martin labs for helpful discussion. A.M. is an HHMI investigator, R.J.D. was an HHMI investigator, and this work was funded by HHMI, the NIH (R01-GM094497 to A.M.), and a gift from Amgen to Caltech. S.N.G. is a Howard Hughes Medical Institute Fellow of the Damon Runyon Cancer Research Foundation, DRG-2342-18. Author Contributions: Conceptualization, E.E.B., S.N.G., R.J.D., and A.M.; Methodology, E.E.B., S.N.G., R.J.D., and A.M; Investigation, E.E.B. and S.N.G.; Writing – Original Draft, E.E.B., S.N.G., and A.M.; Writing – Review & Editing, E.E.B., S.N.G., R.J.D., and A.M; Supervision, R.J.D. and A.M.; Funding Acquisition, R.J.D. and A.M. E.E.B., S.N.G., and A.M. declare no competing interests. R.J.D. is currently Senior Vice President of Global Research at Amgen.

Attached Files

Accepted Version - nihms-1545122.pdf

Supplemental Material - 1-s2.0-S096921261930317X-mmc1.pdf


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