Hsp90-mediated assembly of the 26 S proteasome is involved in major histocompatibility complex class I antigen processing

Taketoshi Yamano, Shusaku Mizukami, Shigeo Murata, Tomoki Chiba, Keiji Tanaka, Heiichiro Udono

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Heat shock protein 90 (hsp90) and the proteasome activator PA28 stimulate major histocompatibility complex (MHC) class I antigen processing. It is unknown whether hsp90 influences the proteasome activity to produce T cell epitopes, although association of PA28 with the 20 S proteasome stimulates the enzyme activity. Here, we show that hsp90 is essential in assembly of the 26 S proteasome and as a result, is involved in epitope production. Addition of recombinant hsp90α to cell lysate enhanced chymotrypsin-like activity of the 26 S proteasome in an ATP-dependent manner as determined by an in-gel hydrolysis assay. We successfully pulled down histidine-tagged hsp90α- and PA28α-induced, newly assembled 26 S proteasomes from the cell extracts for in vitro epitope production assay, and we found these structures to be sensitive to geldanamycin, an hsp90 inhibitor. We found a cleaved epitope unique to the proteasome pulled down by both hsp90α and PA28α, whereas two different epitopes were identified in the hsp90α- and PA28α-pulldowns, respectively. Processing of these respective peptides in vivo was enhanced faithfully by the protein combinations used for the proteasome pulldowns. Inhibition of hsp90 in vivo by geldanamycin partly disrupted the 26 S proteasome structure, consistent with down-regulated MHC class I expression. Our results indicate that hsp90 facilitates MHC class I antigen processing through epitope production in a complex of the 26 S proteasome.

Original languageEnglish
Pages (from-to)28060-28065
Number of pages6
JournalJournal of Biological Chemistry
Volume283
Issue number42
DOIs
Publication statusPublished - Oct 17 2008
Externally publishedYes

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HSP90 Heat-Shock Proteins
Histocompatibility Antigens Class I
Antigen Presentation
Proteasome Endopeptidase Complex
Major Histocompatibility Complex
Processing
Epitopes
Assays
T-Lymphocyte Epitopes
Chymotrypsin
Enzyme activity
Cell Extracts
Recombinant Proteins
Histidine
Hydrolysis
Adenosine Triphosphate
Gels
Association reactions

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Hsp90-mediated assembly of the 26 S proteasome is involved in major histocompatibility complex class I antigen processing. / Yamano, Taketoshi; Mizukami, Shusaku; Murata, Shigeo; Chiba, Tomoki; Tanaka, Keiji; Udono, Heiichiro.

In: Journal of Biological Chemistry, Vol. 283, No. 42, 17.10.2008, p. 28060-28065.

Research output: Contribution to journalArticle

Yamano, Taketoshi ; Mizukami, Shusaku ; Murata, Shigeo ; Chiba, Tomoki ; Tanaka, Keiji ; Udono, Heiichiro. / Hsp90-mediated assembly of the 26 S proteasome is involved in major histocompatibility complex class I antigen processing. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 42. pp. 28060-28065.
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