A novel proteasome interacting protein recruits the deubiquitinating enzyme UCH37 to 26S proteasomes

EMBO J. 2006 Oct 4;25(19):4524-36. doi: 10.1038/sj.emboj.7601338. Epub 2006 Sep 21.

Abstract

The 26S proteasome is a multisubunit protease responsible for regulated proteolysis in eukaryotic cells. It is composed of one catalytic 20S proteasome and two 19S regulatory particles attached on both ends of 20S proteasomes. Here, we describe the identification of Adrm1 as a novel proteasome interacting protein in mammalian cells. Although the overall sequence of Adrm1 has weak homology with the yeast Rpn13, the amino- and carboxyl-terminal regions exhibit significant homology. Therefore, we designated it as hRpn13. hRpn13 interacts with a base subunit Rpn2 via its amino-terminus. The majority of 26S proteasomes contain hRpn13, but a portion of them does not, indicating that hRpn13 is not an integral subunit. Intriguingly, we found that hRpn13 recruits UCH37, a deubiquitinating enzyme known to associate with 26 proteasomes. The carboxyl-terminal regions containing KEKE motifs of both hRpn13 and UCH37 are involved in their physical interaction. Knockdown of hRpn13 caused no obvious proteolytic defect but loss of UCH37 proteins and decrease in deubiquitinating activity of 26S proteasomes. Our results indicate that hRpn13 is essential for the activity of UCH37.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carboxypeptidases
  • Carrier Proteins / metabolism*
  • Conserved Sequence
  • HeLa Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / metabolism*
  • Molecular Sequence Data
  • Proteasome Endopeptidase Complex / metabolism*
  • Protein Binding
  • Protein Processing, Post-Translational
  • Protein Subunits / metabolism
  • RNA, Small Interfering
  • Saccharomyces cerevisiae Proteins / chemistry
  • Sequence Alignment
  • Ubiquitin / metabolism*
  • Ubiquitin Thiolesterase

Substances

  • ADRM1 protein, human
  • Carrier Proteins
  • Intracellular Signaling Peptides and Proteins
  • Membrane Glycoproteins
  • Protein Subunits
  • RNA, Small Interfering
  • Saccharomyces cerevisiae Proteins
  • Ubiquitin
  • Carboxypeptidases
  • UCHL5 protein, human
  • Ubiquitin Thiolesterase
  • Proteasome Endopeptidase Complex
  • ATP dependent 26S protease