20S proteasome-dependent generation of an IEpp89 murine cytomegalovirus-derived H-2Ld epitope from a recombinant protein

Antje Voigt; Ulrike Salzmann; Ulrike Seifert; Margitta Dathe; Andrea Soza; Peter Michael Kloetzel; Ulrike Kuckelkorn

doi:10.1016/j.bbrc.2007.02.006

20S proteasome-dependent generation of an IEpp89 murine cytomegalovirus-derived H-2L^d epitope from a recombinant protein

Antje Voigt, Ulrike Salzmann, Ulrike Seifert, Margitta Dathe, Andrea Soza, Peter Michael Kloetzel^*, Ulrike Kuckelkorn

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

The majority of MHC class I epitopes is generated through the ubiquitin-proteasome system. In the present study, we have analyzed the proteasome-dependent generation of the IE pp89 MCMV-derived H-2L^d epitope by both in vitro and in vivo experiments. As revealed by cytotoxic T-cell assays, the pp89 9mer epitope was generated with high fidelity from the recombinant IE pp89 by 20S proteasomes. In vitro processing showed that the recombinant pp89 was rapidly degraded by 20S proteasomes. Analysis of cell lysates under conditions that allowed detection of polyubiquitinated proteins provided no evidence for the presence of ubiquitin-pp89-conjugates in vivo. These findings suggest a ubiquitin-independent mechanism of proteasomal degradation for pp89.

Original language	English
Pages (from-to)	549-554
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	355
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2007.02.006
State	Published - 2007
Externally published	Yes

Bibliographical note

Funding Information:
We thank I. Drung for technical assistance, Dr. Hengel for supplying antibodies, and Dr. Bohmann for plasmids. This work was supported by a grant of Deutsche Forschungsgemeinschaft SFB421 to P.-M. Kloetzel.

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2007.02.006

Cite this

@article{e22da627db34466cbf643733bab20ab6,

title = "20S proteasome-dependent generation of an IEpp89 murine cytomegalovirus-derived H-2Ld epitope from a recombinant protein",

abstract = "The majority of MHC class I epitopes is generated through the ubiquitin-proteasome system. In the present study, we have analyzed the proteasome-dependent generation of the IE pp89 MCMV-derived H-2Ld epitope by both in vitro and in vivo experiments. As revealed by cytotoxic T-cell assays, the pp89 9mer epitope was generated with high fidelity from the recombinant IE pp89 by 20S proteasomes. In vitro processing showed that the recombinant pp89 was rapidly degraded by 20S proteasomes. Analysis of cell lysates under conditions that allowed detection of polyubiquitinated proteins provided no evidence for the presence of ubiquitin-pp89-conjugates in vivo. These findings suggest a ubiquitin-independent mechanism of proteasomal degradation for pp89.",

keywords = "Proteasome, Ubiquitinylation, Viral antigen processing, pp89",

author = "Antje Voigt and Ulrike Salzmann and Ulrike Seifert and Margitta Dathe and Andrea Soza and Kloetzel, {Peter Michael} and Ulrike Kuckelkorn",

note = "Funding Information: We thank I. Drung for technical assistance, Dr. Hengel for supplying antibodies, and Dr. Bohmann for plasmids. This work was supported by a grant of Deutsche Forschungsgemeinschaft SFB421 to P.-M. Kloetzel.",

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journal = "Biochemical and Biophysical Research Communications",

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T1 - 20S proteasome-dependent generation of an IEpp89 murine cytomegalovirus-derived H-2Ld epitope from a recombinant protein

AU - Voigt, Antje

AU - Salzmann, Ulrike

AU - Seifert, Ulrike

AU - Dathe, Margitta

AU - Soza, Andrea

AU - Kloetzel, Peter Michael

AU - Kuckelkorn, Ulrike

N1 - Funding Information: We thank I. Drung for technical assistance, Dr. Hengel for supplying antibodies, and Dr. Bohmann for plasmids. This work was supported by a grant of Deutsche Forschungsgemeinschaft SFB421 to P.-M. Kloetzel.

PY - 2007/4/6

Y1 - 2007/4/6

N2 - The majority of MHC class I epitopes is generated through the ubiquitin-proteasome system. In the present study, we have analyzed the proteasome-dependent generation of the IE pp89 MCMV-derived H-2Ld epitope by both in vitro and in vivo experiments. As revealed by cytotoxic T-cell assays, the pp89 9mer epitope was generated with high fidelity from the recombinant IE pp89 by 20S proteasomes. In vitro processing showed that the recombinant pp89 was rapidly degraded by 20S proteasomes. Analysis of cell lysates under conditions that allowed detection of polyubiquitinated proteins provided no evidence for the presence of ubiquitin-pp89-conjugates in vivo. These findings suggest a ubiquitin-independent mechanism of proteasomal degradation for pp89.

AB - The majority of MHC class I epitopes is generated through the ubiquitin-proteasome system. In the present study, we have analyzed the proteasome-dependent generation of the IE pp89 MCMV-derived H-2Ld epitope by both in vitro and in vivo experiments. As revealed by cytotoxic T-cell assays, the pp89 9mer epitope was generated with high fidelity from the recombinant IE pp89 by 20S proteasomes. In vitro processing showed that the recombinant pp89 was rapidly degraded by 20S proteasomes. Analysis of cell lysates under conditions that allowed detection of polyubiquitinated proteins provided no evidence for the presence of ubiquitin-pp89-conjugates in vivo. These findings suggest a ubiquitin-independent mechanism of proteasomal degradation for pp89.

KW - Proteasome

KW - Ubiquitinylation

KW - Viral antigen processing

KW - pp89

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DO - 10.1016/j.bbrc.2007.02.006

M3 - Article

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SN - 0006-291X

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EP - 554

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

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