Protein Phosphatase-1 Complex Disassembly by p97 is Initiated through Multivalent Recognition of Catalytic and Regulatory Subunits by the p97 SEP-domain Adapters

Kracht, Matthias; Boom, Johannes van den; Seiler, Jonas; Kröning, Alexander; Kaschani, Farnusch; Kaiser, Markus; Meyer, Hemmo

doi:10.1016/j.jmb.2020.10.001

Cited by 24 publications

(32 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This strongly suggests that SPRTN alone cannot serve as a ubiquitin substrate adapter for p97, despite the fact that SPRTN harbors both a ubiquitin-binding UBZ domain as well as a SHP box motif that could bridge the substrate and p97. This can be explained by the fact that substrate adapters such as Ufd1-Npl4 not only link p97 to the ubiquitylated substrate but also have specific structural features to guide the substrate into the central channel of p97 for unfolding ( 19 , 26 ). Consistent with a role of Ufd1-Npl4 in SPRTN-mediated proteolysis, Ufd1-Npl4 was shown to associate with SPRTN along with p97 in cells ( 14 ).…”

Section: Discussionmentioning

confidence: 99%

Ubiquitin-directed AAA+ ATPase p97/VCP unfolds stable proteins crosslinked to DNA for proteolysis by SPRTN

Kröning

Boom

Kracht

et al. 2022

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Ubiquitin-directed AAA+ ATPase p97/VCP unfolds stable proteins crosslinked to DNA for proteolysis by SPRTN

Kröning

Boom

Kracht

et al. 2022

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

“…p97/Cdc48 is involved in a variety of cellular events. It collaborates with diverse cofactors and adaptors to deal with different types of substrates 1 , 2 , 5 , 11 , 28 , 46 , 47 . Some of these cofactors bind to the C-terminal segment that follows the structurally resolved CTE of p97/Cdc48.…”

Section: Discussionmentioning

confidence: 99%

“…As a master molecular machine in protein quality control, p97/Cdc48 is critical for cellular protein homeostasis 3 , 4 . In complex with a variety of cofactors and adaptors, p97/Cdc48 acts as a segregase or unfoldase that extracts polyubiquitinated proteins from membranes, chromatin, macromolecular complexes, and misfolded protein aggregates for subsequent degradation or remodeling 1 , 3 , 5 – 7 . As such, it regulates diverse cellular processes, including endoplasmic reticulum-associated degradation, mitochondrial-associated degradation, membrane fusion, and DNA replication and repair 1 – 3 .…”

Section: Introductionmentioning

confidence: 99%

Cryo-EM structures of human p97 double hexamer capture potentiated ATPase-competent state

Gao

et al. 2022

Cell Discov

View full text Add to dashboard Cite

The conserved ATPase p97 (Cdc48 in yeast) and adaptors mediate diverse cellular processes through unfolding polyubiquitinated proteins and extracting them from macromolecular assemblies and membranes for disaggregation and degradation. The tandem ATPase domains (D1 and D2) of the p97/Cdc48 hexamer form stacked rings. p97/Cdc48 can unfold substrates by threading them through the central pore. The pore loops critical for substrate unfolding are, however, not well-ordered in substrate-free p97/Cdc48 conformations. How p97/Cdc48 organizes its pore loops for substrate engagement is unclear. Here we show that p97/Cdc48 can form double hexamers (DH) connected through the D2 ring. Cryo-EM structures of p97 DH reveal an ATPase-competent conformation with ordered pore loops. The C-terminal extension (CTE) links neighboring D2s in each hexamer and expands the central pore of the D2 ring. Mutations of Cdc48 CTE abolish substrate unfolding. We propose that the p97/Cdc48 DH captures a potentiated state poised for substrate engagement.

show abstract

“…Disassembly is achieved by threading I3 through the p97 channel which strips I3 (and SDS22) off PP1 to allow PP1 holoenzyme formation with other PP1 subunits (Weith et al, 2018). Notably, I3 targeting relies on multivalent recognition of the SDS22-PP1-I3 (SPI) complex by the p37 adapter (Kracht et al, 2020), and involves an internal recognition site within I3 that is inserted as a loop into the p97 pore to initiate unfolding of I3 (van den Boom et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Structural basis of ubiquitin-independent PP1 complex disassembly by p97

Boom

Meyer

Saibil

2022

Preprint

Self Cite

View full text Add to dashboard Cite

The AAA+ ATPase p97 (also called VCP, or Cdc48 in yeast) unfolds proteins and disassembles protein complexes in a myriad of cellular processes, but how a substrate complex needs to be loaded onto p97 by a dedicated substrate adapter and then disassembled by p97 has not been structurally visualized so far. Here we present cryo-EM structures of p97 in the process of disassembling a protein phosphatase-1 (PP1) complex by stripping off an inhibitory subunit. We show that PP1 and its partners SDS22 and inhibitor-3 (I3) bind to a peripheral N-domain of p97 via a direct contact between SDS22 and a groove in the N-domain. A density consistent with the SHP box of the p37 adapter binds to the same N-domain underneath the PP1 complex, while the p37-UBX domain is found on the adjacent N-domain. I3 is likely represented by three densities. One covers the PP1 catalytic site adjacent to SDS22, another is at the PP1 binding site for the RVXF motif in I3 pointing towards the p97 pore, and the third is a peptide threaded through the central channel of the spiral-shaped p97 hexamer. Our data show how p97 arranges a substrate complex between the N-domain and central channel, and then extracts one component by threading it through the channel to disassemble the complex.

show abstract

Protein Phosphatase-1 Complex Disassembly by p97 is Initiated through Multivalent Recognition of Catalytic and Regulatory Subunits by the p97 SEP-domain Adapters

Cited by 24 publications

References 33 publications

Ubiquitin-directed AAA+ ATPase p97/VCP unfolds stable proteins crosslinked to DNA for proteolysis by SPRTN

Ubiquitin-directed AAA+ ATPase p97/VCP unfolds stable proteins crosslinked to DNA for proteolysis by SPRTN

Cryo-EM structures of human p97 double hexamer capture potentiated ATPase-competent state

Structural basis of ubiquitin-independent PP1 complex disassembly by p97

Contact Info

Product

Resources

About