The Angelman syndrome protein Ube3a/E6AP is required for Golgi acidification and surface protein sialylation

J Neurosci. 2013 Feb 27;33(9):3799-814. doi: 10.1523/JNEUROSCI.1930-11.2013.

Abstract

Angelman syndrome (AS) is a severe disorder of postnatal brain development caused by neuron-specific loss of the HECT (homologous to E6AP carboxy terminus) domain E3 ubiquitin ligase Ube3a/E6AP. The cellular role of Ube3a remains enigmatic despite recent descriptions of synaptic and behavioral deficits in AS mouse models. Although neuron-specific imprinting is thought to limit the disease to the brain, Ube3a is expressed ubiquitously, suggesting a broader role in cellular function. In the current study, we demonstrate a profound structural disruption and cisternal swelling of the Golgi apparatus (GA) in the cortex of AS (UBE3A(m-/p+)) mice. In Ube3a knockdown cell lines and UBE3A(m-/p+) cortical neurons, the GA is severely under-acidified, leading to osmotic swelling. Both in vitro and in vivo, the loss of Ube3a and corresponding elevated pH of the GA is associated with a marked reduction in protein sialylation, a process highly dependent on intralumenal Golgi pH. Altered ion homeostasis of the GA may provide a common cellular pathophysiology underlying the diverse plasticity and neurodevelopmental deficits associated with AS.

Publication types

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

MeSH terms

  • Analysis of Variance
  • Angelman Syndrome / genetics
  • Angelman Syndrome / pathology
  • Animals
  • Animals, Newborn
  • Bacterial Proteins / genetics
  • Cells, Cultured
  • Cerebral Cortex / cytology
  • Cerebral Cortex / ultrastructure*
  • Cytoplasmic Structures / genetics
  • Cytoplasmic Structures / metabolism
  • Cytoplasmic Structures / ultrastructure
  • Disease Models, Animal
  • Embryo, Mammalian
  • Female
  • Gene Expression Regulation / genetics
  • Gene Expression Regulation / physiology
  • Glycine / analogs & derivatives
  • Golgi Apparatus / genetics*
  • Golgi Apparatus / pathology*
  • Golgi Apparatus / ultrastructure
  • Green Fluorescent Proteins / genetics
  • HEK293 Cells
  • Humans
  • Hydrogen-Ion Concentration
  • Lectins / metabolism
  • Luminescent Proteins / genetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microscopy, Electron, Transmission
  • Mutagenesis
  • N-Acetylneuraminic Acid / metabolism*
  • Neurons / metabolism
  • Neurons / ultrastructure
  • Protein Transport / genetics
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Spermine / analogs & derivatives
  • Transfection
  • Ubiquitin-Protein Ligases / deficiency
  • Ubiquitin-Protein Ligases / metabolism*
  • Vesicle-Associated Membrane Protein 2 / genetics
  • Vesicle-Associated Membrane Protein 2 / metabolism
  • Viral Proteins / genetics
  • Viral Proteins / metabolism

Substances

  • Bacterial Proteins
  • Lectins
  • Luminescent Proteins
  • RNA, Small Interfering
  • Vesicle-Associated Membrane Protein 2
  • Viral Proteins
  • yellow fluorescent protein, Bacteria
  • dioctadecylamidoglycylspermine
  • Green Fluorescent Proteins
  • Spermine
  • Ube3a protein, mouse
  • Ubiquitin-Protein Ligases
  • N-Acetylneuraminic Acid
  • Glycine