Invited Speaker 50th Lorne Proteins Conference 2025

Structural basis of proteasomal recognition of a branched ubiquitin chain (114536)

Shang-Te Danny Hsu 1 2 3
  1. Institute of Biological Chemsitry, Academia Sinica, Taipei, Taiwan
  2. Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan
  3. International Institute for Sustainability with Knotted Chiral Meta Matter, Hiroshima University, Hiroshima, Japan

Proteasome-mediated proteolysis is primarily understood through the lens of substrates modified with homotypic K48-linked ubiquitin chains. Recent studies suggest that K11/K48-branched ubiquitination acts as a rapid proteolysis signal, facilitating substrate turnover during cell cycle checkpoints or under protein unfolding stress. Using cryo-electron microscopy single particle reconstruction (cryo-EM SPR), we determined the structures of multiple functional states of the human 26S proteasome in complex with a substrate featuring a K11/K48-branched ubiquitin chain. Our findings reveal a distinct substrate binding groove on the 19S regulatory particle, involving several proteasomal subunits, including Rpn2, which recognizes K48-linked di-ubiquitin through a structural motif similar to that of the well-known ubiquitin receptor Rpn1. The branching point of K11/K48 is located on the proximal ubiquitin, enabling multivalent interactions that likely enhance substrate binding affinity. This mechanism underscores the role of K11/K48-branched ubiquitination in promoting rapid substrate turnover under specific cellular conditions.