Brucella infection inhibits macrophages apoptosis via Nedd4-dependent degradation of calpain2

Vet Microbiol. 2014 Nov 7;174(1-2):195-205. doi: 10.1016/j.vetmic.2014.08.033. Epub 2014 Sep 11.

Abstract

The calcium-dependent protease calpain2 is involved in macrophages apoptosis. Brucella infection-induced up-regulation of intracellular calcium level is an essential factor for the intracellular survival of Brucella within macrophages. Here, we hypothesize that calcium-dependent E3 ubiquitin ligase Nedd4 ubiquitinates calpain2 and inhibits Brucella infection-induced macrophage apoptosis via degradation of calpain2.Our results reveal that Brucella infection induces increases in Nedd4 activity in an intracellular calcium dependent manner. Furthermore, Brucella infection-induced degradation of calpain2 is mediated by Nedd4 ubiquitination of calpain2. Brucella infection-induced calpain2 degradation inhibited macrophages apoptosis. Treatment of Brucella infected macrophages with calcium chelator BAPTA or Nedd4 knock-down decreased Nedd4 activity, prevented calpain2 degradation, and resulted in macrophages apoptosis.

Keywords: Apoptosis; Brucella abortus; Calpain2; Intracellular calcium; Nedd4.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Apoptosis / genetics
  • Apoptosis / physiology*
  • Blotting, Western
  • Brucella / genetics
  • Brucella / pathogenicity*
  • Brucellosis / immunology*
  • Calpain / metabolism
  • Cell Line
  • Densitometry
  • Electrophoresis, Polyacrylamide Gel
  • Endosomal Sorting Complexes Required for Transport / metabolism*
  • Gene Expression Regulation / physiology*
  • Humans
  • Immunoprecipitation
  • In Situ Nick-End Labeling
  • Macrophages / microbiology*
  • Macrophages / physiology
  • Mice
  • Microscopy, Fluorescence
  • Nedd4 Ubiquitin Protein Ligases
  • RNA Interference
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination

Substances

  • Endosomal Sorting Complexes Required for Transport
  • Nedd4 Ubiquitin Protein Ligases
  • Nedd4 protein, human
  • Nedd4l protein, mouse
  • Ubiquitin-Protein Ligases
  • Calpain