Ubiquitin-mediated proteolysis centers in HeLa cells: Indication from studies of an inhibitor of the chymotrypsin-like activity of the proteasome

Cezary Wójcik, Dieter Schroeter, Sherwin Wilk, Jan Lamprecht, Neidhard Paweletz

Research output: Contribution to journalArticle

124 Scopus citations

Abstract

HeLa cells growing in vitro were treated with the peptidyl aldehyde inhibitor of the chymotrypsin-like activity of the proteasome N-benzyloxycarbonyl-ILe- Glu(O-t-butyl)-Ala-leucinal (PSI). Immunefluorescence studies of treated cells revealed the formation of massive perinuclear aggregates rich in ubiquitin and proteasomal antigens, which on the ultrastructural level appeared as perinuclear aggregates of electron-dense material, usually in the vicinity of Golgi cisternae. Histochemical studies disclosed that these cells contained protein-rich perinuclear aggregates detected by amido black staining, while unusual accumulations of lipids, carbohydrates, or nucleic acids were not present. Inhibition of protein synthesis by cycloheximide prevented the formation of aggregates, whereas microtubule disruption by nocodazole induced a dispersion of the aggregates. We hypothesize that aggregates induced by PSI treatment correspond to accumulations of proteasome-substrate complexes in a well-defined region, where the proteolytic processes of the ubiquitin-proteasome pathway seem to be somehow centered. We propose to call this region the proteolysis center.

Original languageEnglish (US)
Pages (from-to)311-318
Number of pages8
JournalEuropean Journal of Cell Biology
Volume71
Issue number3
StatePublished - Nov 1996

Keywords

  • HeLa cells
  • Prosomes
  • Proteasomes
  • Proteolysis center
  • Ubiquitin

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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