Harnessing the ubiquitination machinery to target the degradation of specific cellular proteins

Mol Cell. 2000 Sep;6(3):751-6. doi: 10.1016/s1097-2765(00)00074-5.

Abstract

The functional characterization of a specific gene, or its protein product, often relies on assessing the consequences of its elimination, usually accomplished by gene knockout, ribozyme, antisense, or RNA-mediated interference (RNAi) technologies. The selective degradation of cellular proteins is mediated primarily by the ubiquitin-proteasome pathway. Manipulation of the ubiquitin-dependent proteolytic machinery to eliminate specific gene products at the protein level has been previously attempted with some success in vitro; however, the in vivo efficacy of this approach has not yet been achieved. Here we report successful engineering of the substrate receptor of a major ubiquitin-proteolytic machinery to direct the degradation of otherwise stable cellular proteins both in yeast and in mammalian cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • F-Box Proteins*
  • F-Box-WD Repeat-Containing Protein 7
  • Female
  • GTP-Binding Proteins / metabolism
  • Gene Expression Regulation, Enzymologic
  • Humans
  • Ligases / genetics*
  • Ligases / metabolism
  • Mammals
  • Molecular Biology / methods*
  • Nuclear Proteins / metabolism
  • Osteosarcoma
  • Plasmids
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Retinoblastoma Protein / metabolism*
  • Retinoblastoma-Like Protein p107
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Substrate Specificity / genetics
  • Tumor Cells, Cultured
  • Ubiquitin-Protein Ligases
  • Ubiquitins / metabolism*
  • Uterine Cervical Neoplasms
  • beta-Transducin Repeat-Containing Proteins

Substances

  • BTRC protein, human
  • CDC4 protein, S cerevisiae
  • Cell Cycle Proteins
  • F-Box Proteins
  • F-Box-WD Repeat-Containing Protein 7
  • FBXW7 protein, human
  • Nuclear Proteins
  • Recombinant Fusion Proteins
  • Retinoblastoma Protein
  • Retinoblastoma-Like Protein p107
  • Saccharomyces cerevisiae Proteins
  • Ubiquitins
  • beta-Transducin Repeat-Containing Proteins
  • Ubiquitin-Protein Ligases
  • GTP-Binding Proteins
  • Ligases