Glutamine deamidation and dysfunction of ubiquitin/NEDD8 induced by a bacterial effector family

Science. 2010 Sep 3;329(5996):1215-8. doi: 10.1126/science.1193844. Epub 2010 Aug 5.

Abstract

A family of bacterial effectors including Cif homolog from Burkholderia pseudomallei (CHBP) and Cif from Enteropathogenic Escherichia coli (EPEC) adopt a functionally important papain-like hydrolytic fold. We show here that CHBP was a potent inhibitor of the eukaryotic ubiquitination pathway. CHBP acted as a deamidase that specifically and efficiently deamidated Gln40 in ubiquitin and ubiquitin-like protein NEDD8 both in vitro and during Burkholderia infection. Deamidated ubiquitin was impaired in supporting ubiquitin-chain synthesis. Cif selectively deamidated NEDD8, which abolished the activity of neddylated Cullin-RING ubiquitin ligases (CRLs). Ubiquitination and ubiquitin-dependent degradation of multiple CRL substrates were impaired by Cif in EPEC-infected cells. Mutations of substrate-contacting residues in Cif abolished or attenuated EPEC-induced cytopathic phenotypes of cell cycle arrest and actin stress fiber formation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / metabolism*
  • Bacterial Proteins / metabolism*
  • Burkholderia / pathogenicity
  • Burkholderia pseudomallei / metabolism*
  • Burkholderia pseudomallei / pathogenicity
  • Cell Cycle
  • Cell Line
  • Cullin Proteins / metabolism
  • Enteropathogenic Escherichia coli / metabolism*
  • Enteropathogenic Escherichia coli / pathogenicity
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Glutamine / metabolism*
  • HeLa Cells
  • Humans
  • NEDD8 Protein
  • Point Mutation
  • Stress Fibers / metabolism
  • Transfection
  • Ubiquitin / metabolism*
  • Ubiquitin C / metabolism
  • Ubiquitin-Conjugating Enzymes / metabolism
  • Ubiquitin-Protein Ligases / metabolism
  • Ubiquitination
  • Ubiquitins / metabolism*

Substances

  • Bacterial Proteins
  • Cif protein, E coli
  • Cullin Proteins
  • Escherichia coli Proteins
  • NEDD8 Protein
  • NEDD8 protein, human
  • Ubiquitin
  • Ubiquitin C
  • Ubiquitins
  • Glutamine
  • Ubiquitin-Conjugating Enzymes
  • Ubiquitin-Protein Ligases
  • Amidohydrolases