9T64

Cytoplasmic lattice filament repeat unit with the central FBXW-SKP1 complex

This is a non-PDB format compatible entry.

Summary for 9T64

• Entry DOI: 10.2210/pdb9t64/pdb
• EMDB information: 55604
• Descriptor: KH domain-containing protein 3, Ubiquitin-conjugating enzyme E2 D3, Tubulin alpha-1C chain, ... (14 entities in total)
• Functional Keywords: ubiquitination, complex, filament, unknown function
• Biological source: Mus musculus (house mouse); More
• Total number of polymer chains: 69
• Total formula weight: 4531834.58

Authors

Singh, K.,Harasimov, K.,Carter, A.P. (deposition date: 2025-11-06, release date: 2026-08-12, Last modification date: 2026-08-26)

Primary citation

Singh, K.,Harasimov, K.,Niakan, K.K.,Carter, A.P.
In-situ cryo-ET of mouse embryos reveals cytoplasmic lattices contain ubiquitin-charged E2-E3 ligase assemblies.
Embo J., 2026

PubMed Abstract: Cytoplasmic lattices (CPLs) are filamentous assemblies essential for mammalian embryonic development. They are known to regulate organelle organization, spindle assembly, and protein homeostasis, but their molecular functions remain unclear. Here, we develop a strategy combining cryo-focused ion beam milling and cryo-electron tomography to resolve macromolecular complexes directly in mammalian embryos. Using this approach, we determine the in situ structure of cytoplasmic lattices within 6/8-cell mouse embryos at ~4.7 Å resolution. CPL filaments are built from multiple copies of at least fourteen proteins arranged into a ~4.5 MDa repeating unit. The repeat contains a central cavity that is open at the back and lined with multiple FBXW-SKP1 complexes and three modules, each containing the E2 ubiquitin-conjugating enzyme UBE2D and the E3 ligase UHRF1. We resolve two CPL states: one is consistent with a ubiquitin-charged UBE2D, where ubiquitin is held in an open, inactive conformation by binding the scaffold protein PADI6; the second lacks discernible ubiquitin density and shows structural changes compatible with ubiquitin becoming available for transfer. Our findings support a model in which CPLs function as large ubiquitin ligase assemblies during early embryonic development.

PubMed: 42575997
DOI: 10.1038/s44318-026-00895-w

Experimental method

ELECTRON MICROSCOPY (7.3 Å)