Diverse fate of ubiquitin chain moieties: the proximal is degraded with the target, and the distal protects the proximal from removal and recycles

Hao Sun, Sachitanand M. Mali, Sumeet K. Singh, Roman Meledin, Ashraf Brik, Yong Tae Kwon, Yelena Kravtsova-Ivantsiv, Beatrice Bercovich, Aaron Ciechanover

Research output: Contribution to journalArticlepeer-review

43 Scopus citations

Abstract

One of the enigmas in the ubiquitin (Ub) field is the requirement for a poly-Ub chain as a proteasomal targeting signal. The canonical chain appears to be longer than the distance between the two Ub-binding proteasomal receptors. Furthermore, genetic manipulation has shown that one receptor subunit is sufficient, which suggests that a single Ub can serve as a degradation signal. To shed light on this mystery, we chemically synthesized tetra-Ub, di-Ub (K 48 -based), and mono-Ub adducts of HA-α-globin, where the distal or proximal Ub moieties were tagged differentially with either Myc or Flag. When incubated in a crude cell extract, the distal Ub moiety in the tetra-Ub adduct was mostly removed by deubiquitinating enzymes (DUBs) and reconjugated to other substrates in the extract. In contrast, the proximal moiety was most likely degraded with the substrate. The efficacy of degradation was proportionate to the chain length; while tetra-Ub globin was an efficient substrate, with mono-Ub globin, we observed rapid removal of the Ub moiety with almost no degradation of the free globin. Taken together, these findings suggest that the proximal moieties are necessary for securing the association of the substrate with the proteasome along the proteolytic process, whereas the distal moieties are important in protecting the proximal moieties from premature deubiquitination. Interestingly, when the same experiment was carried out using purified 26S proteasome, mono- and tetra-Ub globin were similarly degraded, highlighting the roles of the entire repertoire of cellular DUBs in regulating the degradation of proteasomal substrates.

Original languageEnglish
Pages (from-to)7805-7812
Number of pages8
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number16
DOIs
StatePublished - 16 Apr 2019
Externally publishedYes

Keywords

  • 26S proteasome
  • Chemical synthesis
  • Degradation
  • Deubiquitination
  • Protein
  • Ubiquitination

ASJC Scopus subject areas

  • General

Fingerprint

Dive into the research topics of 'Diverse fate of ubiquitin chain moieties: the proximal is degraded with the target, and the distal protects the proximal from removal and recycles'. Together they form a unique fingerprint.

Cite this