Minimal length requirement for proteasomal degradation of ubiquitin-dependent substrates

FASEB J. 2009 Jan;23(1):123-33. doi: 10.1096/fj.08-115055. Epub 2008 Sep 16.

Abstract

An erroneous transcriptional process, known as molecular misreading, gives rise to an alternative transcript of the ubiquitin B (UBB) gene. This transcript encodes the protein UBB(+1), which comprises a ubiquitin moiety and a 19-aa C-terminal extension. UBB(+1) is found in affected neurons in neurodegenerative diseases and behaves as an atypical ubiquitin fusion degradation (UFD) proteasome substrate that is poorly degraded and impedes the ubiquitin/proteasome system. Here, we show that the limited length of UBB(+1) is responsible for its inefficient degradation and inhibitory activity. Designed UFD substrates with an equally short 19-aa or a 20-aa C-terminal extension were also poorly degraded and had a general inhibitory activity on the ubiquitin/proteasome system in two unrelated cell lines. Extending the polypeptide to 25 aa sufficed to convert the protein into an efficiently degraded proteasome substrate that lacked inhibitory activity. A similar length dependency was found for degradation of two UFD substrates in Saccharomyces cerevisiae, which suggests that the mechanisms underlying this length constraint are highly conserved. Extending UBB(+1) also converted this protein into an efficient substrate of the proteasome. These observations provide an explanation for the accumulation of UBB(+1) in neurodegenerative disorders and offers new insights into the physical constraints determining proteasomal degradation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Line, Tumor
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Proteasome Endopeptidase Complex / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Substrate Specificity
  • Transcription, Genetic
  • Ubiquitin / genetics*
  • Ubiquitin / metabolism*

Substances

  • Ubiquitin
  • Proteasome Endopeptidase Complex