Mechanism of proteasome gate modulation by assembly chaperones Pba1 and Pba2

J Biol Chem. 2022 May;298(5):101906. doi: 10.1016/j.jbc.2022.101906. Epub 2022 Apr 6.

Abstract

The active sites of the proteasome are housed within its central core particle (CP), a barrel-shaped chamber of four stacked heptameric rings, and access of substrates to the CP interior is mediated by gates at either axial end. These gates are constitutively closed and may be opened by the regulatory particle (RP), which binds the CP and facilitates substrate degradation. We recently showed that the heterodimeric CP assembly chaperones Pba1/2 also mediate gate opening through an unexpected structural arrangement that facilitates the insertion of the N terminus of Pba1 into the CP interior; however, the full mechanism of Pba1/2-mediated gate opening is unclear. Here, we report a detailed analysis of CP gate modulation by Pba1/2. The clustering of key residues at the interface between neighboring α-subunits is a critical feature of RP-mediated gate opening, and we find that Pba1/2 recapitulate this strategy. Unlike RP, which inserts at six α-subunit interfaces, Pba1/2 insert at only two α-subunit interfaces. Nevertheless, Pba1/2 are able to regulate six of the seven interfacial clusters, largely through direct interactions. The N terminus of Pba1 also physically interacts with the center of the gate, disrupting the intersubunit contacts that maintain the closed state. This novel mechanism of gate modulation appears to be unique to Pba1/2 and therefore likely occurs only during proteasome assembly. Our data suggest that release of Pba1/2 at the conclusion of assembly is what allows the nascent CP to assume its mature gate conformation, which is primarily closed, until activated by RP.

Keywords: HbYX; Pba1; Pba2; core particle; gate; proteasome.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cytoplasm / metabolism
  • Molecular Chaperones / metabolism
  • Proteasome Endopeptidase Complex* / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins* / metabolism

Substances

  • Molecular Chaperones
  • Pba1 protein, S cerevisiae
  • Pba2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Proteasome Endopeptidase Complex