Document Detail


Visualization of the ER-to-cytosol dislocation reaction of a type I membrane protein.
MedLine Citation:
PMID:  11867532     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The human cytomegalovirus gene products US2 and US11 induce proteasomal degradation of MHC class I heavy chains. We have generated an enhanced green fluorescent protein-class I heavy chain (EGFP-HC) chimeric molecule to study its dislocation and degradation in US2- and US11-expressing cells. The EGFP-HC fusion is stable in control cells, but is degraded rapidly in US2- or US11-expressing cells. Proteasome inhibitors induce in a time-dependent manner the accumulation of EGFP-HC molecules in US2- and US11-expressing cells, as assessed biochemically and by cytofluorimetry of intact cells. Pulse-chase analysis and subcellular fractionation show that EGFP-HC proteins are dislocated from the endoplasmic reticulum and can be recovered as deglycosylated fluorescent intermediates in the cytosol. These results raise the possibility that dislocation of glycoproteins from the ER may not require their full unfolding.
Authors:
Edda Fiebiger; Craig Story; Hidde L Ploegh; Domenico Tortorella
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The EMBO journal     Volume:  21     ISSN:  0261-4189     ISO Abbreviation:  EMBO J.     Publication Date:  2002 Mar 
Date Detail:
Created Date:  2002-02-27     Completed Date:  2002-05-14     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  8208664     Medline TA:  EMBO J     Country:  England    
Other Details:
Languages:  eng     Pagination:  1041-53     Citation Subset:  IM    
Affiliation:
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Armenise Building, Boston, MA 02115, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Astrocytoma / pathology
Brain Neoplasms / pathology
Cysteine Endopeptidases / metabolism
Cytomegalovirus / genetics
Cytosol / metabolism*
Endoplasmic Reticulum / metabolism*
Flow Cytometry
Genes, Synthetic
Glycosylation
Green Fluorescent Proteins
H-2 Antigens / genetics,  metabolism*
HLA-A2 Antigen / genetics,  metabolism*
Humans
Luminescent Proteins / genetics,  metabolism
Membrane Glycoproteins / metabolism*
Mice
Multienzyme Complexes / metabolism
Peptide Fragments / genetics,  metabolism
Proteasome Endopeptidase Complex
Protein Folding
Protein Processing, Post-Translational
Protein Sorting Signals / genetics,  physiology
Protein Transport*
RNA-Binding Proteins / genetics,  metabolism
Recombinant Fusion Proteins / genetics,  metabolism
Subcellular Fractions / chemistry
Transfection
Tumor Cells, Cultured
Viral Envelope Proteins / genetics,  metabolism
Viral Proteins / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
5R37-AI33456/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/H-2 Antigens; 0/H-2Kb protein, mouse; 0/HLA-A2 Antigen; 0/Luminescent Proteins; 0/Membrane Glycoproteins; 0/Multienzyme Complexes; 0/Peptide Fragments; 0/Protein Sorting Signals; 0/RNA-Binding Proteins; 0/Recombinant Fusion Proteins; 0/US11 protein, herpesvirus; 0/US2 protein, Varicellovirus; 0/Viral Envelope Proteins; 0/Viral Proteins; 147336-22-9/Green Fluorescent Proteins; EC 3.4.22.-/Cysteine Endopeptidases; EC 3.4.25.1/Proteasome Endopeptidase Complex
Comments/Corrections

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