Base-CP proteasome can serve as a platform for stepwise lid formation

Zanlin Yu, Nurit Livnat-Levanon, Oded Kleifeld, Wissam Mansour, Mark A. Nakasone, Carlos A. Castaneda, Emma K. Dixon, David Fushman, Noa Reis, Elah Pick, Michael H. Glickman

Research output: Research - peer-reviewArticle

  • 9 Citations

Abstract

26S proteasome, a major regulatory protease in eukaryotes, consists of a 20S proteolytic core particle (CP) capped by a 19S regulatory particle (RP). The 19S RP is divisible into base and lid sub-complexes. Even within the lid, subunits have been demarcated into two modules: module 1 (Rpn5, Rpn6, Rpn8, Rpn9 and Rpn11), which interacts with both CP and base sub-complexes and module 2 (Rpn3, Rpn7, Rpn12 and Rpn15) that is attached mainly to module 1. We now show that suppression of RPN11 expression halted lid assembly yet enabled the base and 20S CP to pre-assemble and form a base-CP. A key role for Regulatory particle non-ATPase 11 (Rpn11) in bridging lid module 1 and module 2 subunits together is inferred from observing defective proteasomes in rpn11-m1, a mutant expressing a truncated form of Rpn11 and displaying mitochondrial phenotypes. An incomplete lid made up of five module 1 subunits attached to base-CP was identified in proteasomes isolated from this mutant. Re-introducing the C-terminal portion of Rpn11 enabled recruitment of missing module 2 subunits. In vitro, module 1 was reconstituted stepwise, initiated by Rpn11-Rpn8 heterodimerization. Upon recruitment of Rpn6, the module 1 intermediate was competent to lock into base-CP and reconstitute an incomplete 26S proteasome. Thus, base-CP can serve as a platform for gradual incorporation of lid, along a proteasome assembly pathway. Identification of proteasome intermediates and reconstitution of minimal functional units should clarify aspects of the inner workings of this machine and how multiple catalytic processes are synchronized within the 26S proteasome holoenzymes.

LanguageEnglish (US)
Article numbere00194
JournalBioscience Reports
Volume35
DOIs
StatePublished - 2015
Externally publishedYes

Fingerprint

Proteasome Endopeptidase Complex
ATP dependent 26S protease
Holoenzymes
Eukaryota
Peptide Hydrolases
Phenotype
In Vitro Techniques

Keywords

  • 19S regulatory particle
  • 20S core particle
  • 26S proteasome
  • Base
  • Lid
  • MPN
  • PCI
  • rpn11-m1

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Yu, Z., Livnat-Levanon, N., Kleifeld, O., Mansour, W., Nakasone, M. A., Castaneda, C. A., ... Glickman, M. H. (2015). Base-CP proteasome can serve as a platform for stepwise lid formation. Bioscience Reports, 35, [e00194]. DOI: 10.1042/BSR20140173

Base-CP proteasome can serve as a platform for stepwise lid formation. / Yu, Zanlin; Livnat-Levanon, Nurit; Kleifeld, Oded; Mansour, Wissam; Nakasone, Mark A.; Castaneda, Carlos A.; Dixon, Emma K.; Fushman, David; Reis, Noa; Pick, Elah; Glickman, Michael H.

In: Bioscience Reports, Vol. 35, e00194, 2015.

Research output: Research - peer-reviewArticle

Yu, Z, Livnat-Levanon, N, Kleifeld, O, Mansour, W, Nakasone, MA, Castaneda, CA, Dixon, EK, Fushman, D, Reis, N, Pick, E & Glickman, MH 2015, 'Base-CP proteasome can serve as a platform for stepwise lid formation' Bioscience Reports, vol 35, e00194. DOI: 10.1042/BSR20140173
Yu Z, Livnat-Levanon N, Kleifeld O, Mansour W, Nakasone MA, Castaneda CA et al. Base-CP proteasome can serve as a platform for stepwise lid formation. Bioscience Reports. 2015;35. e00194. Available from, DOI: 10.1042/BSR20140173
Yu, Zanlin ; Livnat-Levanon, Nurit ; Kleifeld, Oded ; Mansour, Wissam ; Nakasone, Mark A. ; Castaneda, Carlos A. ; Dixon, Emma K. ; Fushman, David ; Reis, Noa ; Pick, Elah ; Glickman, Michael H./ Base-CP proteasome can serve as a platform for stepwise lid formation. In: Bioscience Reports. 2015 ; Vol. 35.
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