Structural basis for the recognition of N-end rule substrates by the UBR box of ubiquitin ligases

Nat Struct Mol Biol. 2010 Oct;17(10):1175-81. doi: 10.1038/nsmb.1907. Epub 2010 Sep 12.

Abstract

The N-end rule pathway is a regulated proteolytic system that targets proteins containing destabilizing N-terminal residues (N-degrons) for ubiquitination and proteasomal degradation in eukaryotes. The N-degrons of type 1 substrates contain an N-terminal basic residue that is recognized by the UBR box domain of the E3 ubiquitin ligase UBR1. We describe structures of the UBR box of Saccharomyces cerevisiae UBR1 alone and in complex with N-degron peptides, including that of the cohesin subunit Scc1, which is cleaved and targeted for degradation at the metaphase-anaphase transition. The structures reveal a previously unknown protein fold that is stabilized by a novel binuclear zinc center. N-terminal arginine, lysine or histidine side chains of the N-degron are coordinated in a multispecific binding pocket. Unexpectedly, the structures together with our in vitro biochemical and in vivo pulse-chase analyses reveal a previously unknown modulation of binding specificity by the residue at position 2 of the N-degron.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Arginine / chemistry
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / metabolism
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / metabolism
  • Crystallography, X-Ray
  • Histidine / chemistry
  • Hydrophobic and Hydrophilic Interactions
  • Lysine / chemistry
  • Models, Molecular
  • Molecular Sequence Data
  • Oligopeptides / chemistry
  • Oligopeptides / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein Interaction Mapping
  • Protein Processing, Post-Translational
  • Protein Structure, Tertiary
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Structure-Activity Relationship
  • Substrate Specificity
  • Ubiquitin / metabolism
  • Ubiquitin-Protein Ligases / chemistry
  • Ubiquitin-Protein Ligases / metabolism*
  • Zinc Fingers / physiology

Substances

  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • MCD1 protein, S cerevisiae
  • Oligopeptides
  • Saccharomyces cerevisiae Proteins
  • Ubiquitin
  • Histidine
  • Arginine
  • UBR1 protein, S cerevisiae
  • Ubiquitin-Protein Ligases
  • Lysine

Associated data

  • PDB/3NIH
  • PDB/3NII
  • PDB/3NIJ
  • PDB/3NIK
  • PDB/3NIL
  • PDB/3NIM
  • PDB/3NIN
  • PDB/3NIS
  • PDB/3NIT