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Cryo-electron microscopy structures of VCP/p97 reveal a new mechanism of oligomerization regulation

ABSTRACT: Summary VCP/p97 is an evolutionarily conserved AAA+ ATPase important for cellular homeostasis. Previous studies suggest that VCP predominantly exists as a homohexamer. Here, we performed structural and biochemical characterization of VCP dodecamer, an understudied state of VCP. The structure revealed an apo nucleotide status that has rarely been captured, a tail-to-tail assembly of two hexamers, and the up-elevated N-terminal domains akin to that seen in the ATP-bound hexamer. Further analyses elucidated a nucleotide status-dependent dodecamerization mechanism, where nucleotide dissociation from the D2 AAA domains induces and promotes VCP dodecamerization. In contrast, nucleotide-free D1 AAA domains are associated with the up-rotation of N-terminal domains, which may prime D1 for ATP binding. These results therefore reveal new nucleotide status-dictated intra- and interhexamer conformational changes and suggest that modulation of D2 domain nucleotide occupancy may serve as a mechanism in controlling VCP oligomeric states. Graphical abstract Highlights • A cryo-EM structure of VCP dodecamer purified with mammalian cells is resolved• The dodecamer structure reveals an apo nucleotide status• A VCP dodecamerization mechanism has been elucidated• Loss of nucleotide from D2 promotes VCP dodecamer formation Biological sciences; Biochemistry; Structural biology

SUBMITTER: Yu G

PROVIDER: S-EPMC8571493 | biostudies-literature |

REPOSITORIES: biostudies-literature

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