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PMID: 9799224 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Subcellular distribution of proteasomes implicates a major location of protein degradation in the nuclear envelope-ER network in yeast.

The EMBO journal ·Vol. 17 ·No. 21 ·1998-11-02 ·Pages 6144-54

Enenkel C, Lehmann A, Kloetzel PM

Abstract

26S proteasomes are the key enzyme complexes responsible for selective turnover of short-lived and misfolded proteins. Based on the assumption that they are dispersed over the nucleoplasm and cytoplasm in all eukaryotic cells, we wanted to determine the subcellular distribution of 26S proteasomes in living yeast cells. For this purpose, we generated yeast strains that express functional green fluorescent protein (GFP) fusions of proteasomal subunits. An alpha subunit of the proteolytically active 20S core complex of the 26S proteasome, Pre6/YOL038w, as well as an ATPase-type subunit of the regulatory 19S cap complex, Cim5/YOL145w, were tagged with GFP. Both chimeras were shown to be incorporated completely into active 26S proteasomes. Microscopic analysis revealed that GFP-labelled 20S as well as 19S subunits are accumulated mainly in the nuclear envelope (NE)-endoplasmic reticulum (ER) network in yeast. These findings were supported by the co-localization and co-enrichment of 26S proteasomes with NE-ER marker proteins. A major location of proteasomal peptide cleavage activity was visualized in the NE-ER network, indicating that proteasomal degradation takes place mainly in this subcellular compartment in yeast.

MeSH Terms
Adenosine Triphosphatases Biomarkers/analysis DNA-Binding Proteins/metabolism Endopeptidases/metabolism Endoplasmic Reticulum/enzymology,metabolism Fungal Proteins/metabolism Green Fluorescent Proteins Luminescent Proteins Microscopy, Confocal Nuclear Envelope/enzymology,metabolism Peptide Hydrolases/metabolism Proteasome Endopeptidase Complex Recombinant Proteins/genetics Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins
Chemicals
Biomarkers DNA-Binding Proteins Fungal Proteins Luminescent Proteins Recombinant Proteins Saccharomyces cerevisiae Proteins Green Fluorescent Proteins Endopeptidases Peptide Hydrolases Proteasome Endopeptidase Complex ATP dependent 26S protease Adenosine Triphosphatases RPT1 protein, S cerevisiae
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Enenkel C
Institut für Biochemie, Humboldt Universität, Universitätsklinikum Charité, Monbijoustrasse 2, D-10117 Berlin, Germany. enenkel@comcom.rz.charite.hu-berlin.de
Lehmann A
Kloetzel P M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.21.6144
Email
enenkel@comcom.rz.charite.hu-berlin.de
Published
1998-11-02
Pages
6144-54
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170941
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.21.6144 PMC Full Text View journal details
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