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TitleMetabolically regulated proteasome supramolecular organization in situ.
Journal, issue, pagesCell, Vol. 189, Issue 4, Page 1153-1169.e16, Year 2026
Publish dateFeb 19, 2026
AuthorsXiaomeng Tang / Lu Qu / Florian Wilfling / Florian Beck / Oliver P Ernst / Brenda A Schulman / Wolfgang Baumeister / Cordula Enenkel /
PubMed AbstractMany proteins localize in membraneless organelles. However, understanding the steps along membraneless organelle formation-and the structural impact on granule constituents-has been hindered by ...Many proteins localize in membraneless organelles. However, understanding the steps along membraneless organelle formation-and the structural impact on granule constituents-has been hindered by limited resolution of intracellular data. We address these challenges through in situ cryo-electron tomography (cryo-ET) along with formation of yeast proteasome storage granules (PSGs). During the transition from proliferation to quiescence, doubly capped 26S proteasomes arrested in an inactive state arrange into ∼7.5 MDa trimeric units, dispersed in the nucleoplasm and congregated along the nuclear envelope near the nuclear pore. 9-Å-resolution cryo-ET structures reveal that cytoplasmic PSGs formed in various energy-limiting conditions are paracrystalline arrays of bundled fibers, assembled from stacking of proteasome trimers. The paracrystalline arrangement maintains a pool of fully assembled inactive 26S proteasomes that are released in energy-rich conditions. Overall, our data reveal structural steps along the assembly of an intracellular membraneless organelle in situ and quinary structure formation controlling a major eukaryotic regulatory machine.
External linksCell / PubMed:41605212
MethodsEM (tomography) / EM (subtomogram averaging)
Resolution9.1 - 27.4 Å
Structure data

EMDB-60949: In situ cryo-electron tomogram of 4days WT cytoplasm 1
Method: EM (tomography)

EMDB-60950: In situ cryo-electron tomogram of 4days WT nucleus (PSG at NE)
Method: EM (tomography)

EMDB-60951: In situ cryo-electron tomogram of 4days WT nucleus (trimer close to NE)
Method: EM (tomography)

EMDB-60952: In situ cryo-electron tomogram of 4days rpn13null nucleus
Method: EM (tomography)

EMDB-60953: In situ cryo-electron tomogram of 1day WT nucleus
Method: EM (tomography)

EMDB-60955: In situ cryo-electron tomogram of 4days rpn9deltaN nucleus
Method: EM (tomography)

EMDB-60957: In situ cryo-electron tomogram of 4days WT cell
Method: EM (tomography)

60960

9iwr

EMDB-60960, PDB-9iwr:
26S proteasome trimer
Method: EM (subtomogram averaging) / Resolution: 9.1 Å

EMDB-60961: 26S proteasome trimer close to nuclear envelope
Method: EM (subtomogram averaging) / Resolution: 27.4 Å

EMDB-64627: In situ cryo-electron tomogram of 4days rpn9 surface mutant nucleus
Method: EM (tomography)

EMDB-64628: In situ cryo-electron tomogram of 18h nucleus
Method: EM (tomography)

EMDB-64629: In situ cryo-electron tomogram of 4days WT cytoplasm 3
Method: EM (tomography)

EMDB-64630: In situ cryo-electron tomogram of 4days glucose 1h WT nucleus
Method: EM (tomography)

EMDB-64631: In situ cryo-electron tomogram of 4days glucose control WT nucleus
Method: EM (tomography)

EMDB-64632: In situ cryo-electron tomogram of SA 1day WT cytoplasm 1
Method: EM (tomography)

EMDB-64633: In situ cryo-electron tomogram of SA 1day WT cytoplasm 2
Method: EM (tomography)

EMDB-64634: In situ cryo-electron tomogram of 4days mlp1delta mlp2delta nucleus
Method: EM (tomography)

EMDB-64635: In vitro cryo-electron tomogram of 4days WT purified
Method: EM (tomography)

EMDB-64636: In situ cryo-electron tomogram of 4days rpn9deltaN nucleus
Method: EM (tomography)

Source
  • saccharomyces cerevisiae s288c (yeast)
KeywordsHYDROLASE / proteasome / trimer / complex / PSG / paracrystalline

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