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Related Experiment Videos

Proteasome activation by REG molecules lacking homolog-specific inserts

Z Zhang1, C Realini, A Clawson

  • 1Department of Biochemistry, University of Utah, Salt Lake City, Utah 84132, USA.

The Journal of Biological Chemistry
|May 23, 1998
PubMed
Summary

The 11 S REG (regulator of proteasome activity) subunits alpha and gamma, but not beta, are not essential for proteasome activation. REGbeta inserts are crucial for optimal proteasome binding and activation.

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Area of Science:

  • * Molecular biology
  • * Biochemistry
  • * Cell biology

Background:

  • * Eukaryotic proteasome activity is enhanced by the 11 S REG (PA28) complex.
  • * REG subunits (alpha, beta, gamma) contain unique sequence regions termed homolog-specific inserts.
  • * These inserts are hypothesized to influence REG-proteasome interactions and cellular localization.

Purpose of the Study:

  • * To investigate the role of homolog-specific inserts in REG subunits (alpha, beta, gamma) in proteasome activation and oligomerization.
  • * To determine the functional significance of the REGbeta insert in proteasome binding and activation.

Main Methods:

  • * Generation of mutant REG subunits (REGalphaDeltai, REGbetaDeltai, REGgammaDeltai) by deleting homolog-specific inserts.
  • * Purification of recombinant mutant and wild-type REG proteins.

Related Experiment Videos

  • * Assays to evaluate proteasome activation and hetero-oligomer formation using human red cell proteasomes.

    • Main Results:

    • * REGalphaDeltai and REGgammaDeltai formed heptamers and activated proteasomes similarly to wild-type proteins.
    • * REGbetaDeltai showed reduced proteasome activation at low concentrations compared to wild-type REGbeta.
    • * Deletion of the REGbeta insert, but not the REGalpha insert, impaired hetero-oligomer binding and activation of proteasomes.

    Conclusions:

    • * REGalpha and REGgamma inserts are dispensable for proteasome binding and activation.
    • * The REGbeta insert is important for efficient binding of REG subunits and their complexes to proteasomes.
    • * REG inserts are not essential for proteasome activation but may play a role in cellular localization of REG-proteasome complexes.