Participation of the proteasomal lid subunit Rpn11 in mitochondrial morphology and function is mapped to a distinct C-terminal domain

Biochem J. 2004 Jul 1;381(Pt 1):275-85. doi: 10.1042/BJ20040008.

Abstract

Substrates destined for degradation by the 26 S proteasome are labelled with polyubiquitin chains. Rpn11/Mpr1, situated in the lid subcomplex, partakes in the processing of these chains or in their removal from substrates bound to the proteasome. Rpn11 also plays a role in maintaining mitochondrial integrity, tubular structure and proper function. The recent finding that Rpn11 participates in proteasome-associated deubiquitination focuses interest on the MPN+ (Mpr1, Pad1, N-terminal)/JAMM (JAB1/MPN/Mov34) metalloprotease site in its N-terminal domain. However, Rpn11 damaged at its C-terminus (the mpr1-1 mutant) causes pleiotropic effects, including proteasome instability and mitochondrial morphology defects, resulting in both proteolysis and respiratory malfunctions. We find that overexpression of WT (wild-type) RPN8, encoding a paralogous subunit that does not contain the catalytic MPN+ motif, corrects proteasome conformations and rescues cell cycle phenotypes, but is unable to correct defects in the mitochondrial tubular system or respiratory malfunctions associated with the mpr1-1 mutation. Transforming mpr1-1 with various RPN8-RPN11 chimaeras or with other rpn11 mutants reveals that a WT C-terminal region of Rpn11 is necessary, and more surprisingly sufficient, to rescue the mpr1-1 mitochondrial phenotype. Interestingly, single-site mutants in the catalytic MPN+ motif at the N-terminus of Rpn11 lead to reduced proteasome-dependent deubiquitination connected with proteolysis defects. Nevertheless, these rpn11 mutants suppress the mitochondrial phenotypes associated with mpr1-1 by intragene complementation. Together, these results point to a unique role for the C-terminal region of Rpn11 in mitochondrial maintenance that may be independent of its role in proteasome-associated deubiquitination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / deficiency
  • Acetyltransferases / genetics
  • Acetyltransferases / physiology
  • Carboxy-Lyases / genetics
  • Carboxy-Lyases / physiology
  • Catalytic Domain / genetics
  • Catalytic Domain / physiology
  • Endopeptidases / deficiency
  • Endopeptidases / genetics
  • Endopeptidases / physiology*
  • Genetic Complementation Test
  • Mitochondria / physiology*
  • Mitochondria / ultrastructure*
  • Peptide Hydrolases / chemistry*
  • Peptide Hydrolases / physiology
  • Peptide Mapping / methods
  • Peptides / genetics
  • Peptides / physiology
  • Phenotype
  • Proteasome Endopeptidase Complex / deficiency
  • Proteasome Endopeptidase Complex / genetics
  • Proteasome Endopeptidase Complex / physiology*
  • Protein Structure, Tertiary / genetics
  • Protein Structure, Tertiary / physiology
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / physiology*
  • Ubiquitin / metabolism

Substances

  • Peptides
  • RPN11 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Ubiquitin
  • Acetyltransferases
  • Mpr1 protein, S cerevisiae
  • Endopeptidases
  • Peptide Hydrolases
  • Proteasome Endopeptidase Complex
  • Rpn8 protein, S cerevisiae
  • ATP dependent 26S protease
  • Carboxy-Lyases
  • phenylacrylic acid decarboxylase