Hypoxia inducible NOD2 interacts with 3-O-sulfogalactoceramide and regulates vesicular homeostasis

Alexey A. Nabatov, Pantelis Hatzis, Kasper M A Rouschop, Paul Van Diest, Marc Vooijs

Research output: Contribution to journalArticle

Abstract

Background Oxygen sensing in mammalian cells is a conserved signaling pathway regulated by hypoxia inducible factor type 1 (HIF-1). Inadequate oxygen supply (hypoxia) is common to many pathological disorders where autophagy plays an import role. The aim of this study was the identification and characterization of novel HIF-1 target genes that promote autophagy during hypoxia. Methods Whole genome Chromatin Immune Precipitation from hypoxic HeLa cells was used to identify novel HIF-1 target genes. Hypoxia induced expression and transcription regulation was studied in wild type and HIF-deficient cells. siRNA silencing of candidate genes was used to establish their role during autophagy. Recombinant protein was used for screening immobilized glycosylated lipids to identify potential ligands. Results We identified the Nucleotide Oligomerization Domain 2 (NOD2/CARD15) as a novel HIF-1 target and 3-O-sulfo-galactoceramide (sulfatide) and Mycobacterium sp. specific sulfolipid-1 as the first NOD2 ligands that both compete for binding to NOD2. Loss of NOD2 function impaired autophagy upstream of the autophagy inhibitor chloroquine by reducing the number of acidic vesicles. Inhibition of sulfatide synthesis elicited defects in autophagy similar to the NOD2 loss of function but did not influence NOD2-mediated NF-kB signaling. Conclusions Our findings suggest that the interaction of NOD2 with sulfatide may mediate the balance between autophagy and inflammation in hypoxic cells. General significance These findings may lead to a better understanding of complex inflammatory pathologies like Crohn's disease and tuberculosis where both NOD2 and hypoxia are implicated.

Original languageEnglish (US)
Pages (from-to)5277-5286
Number of pages10
JournalBiochimica et Biophysica Acta - General Subjects
Volume1830
Issue number11
DOIs
StatePublished - 2013
Externally publishedYes

Fingerprint

Hypoxia-Inducible Factor 1
Autophagy
Homeostasis
Genes
Cells
Ligands
Oxygen supply
Oligomerization
NF-kappa B
Chloroquine
Pathology
Transcription
Recombinant Proteins
Small Interfering RNA
Chromatin
Screening
Oxygen
Nucleotides
Lipids
Defects

Keywords

  • Autophagy
  • Hypoxia
  • NOD2
  • Sulfatide
  • V-ATPase
  • Vesicle

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Hypoxia inducible NOD2 interacts with 3-O-sulfogalactoceramide and regulates vesicular homeostasis. / Nabatov, Alexey A.; Hatzis, Pantelis; Rouschop, Kasper M A; Van Diest, Paul; Vooijs, Marc.

In: Biochimica et Biophysica Acta - General Subjects, Vol. 1830, No. 11, 2013, p. 5277-5286.

Research output: Contribution to journalArticle

Nabatov, Alexey A. ; Hatzis, Pantelis ; Rouschop, Kasper M A ; Van Diest, Paul ; Vooijs, Marc. / Hypoxia inducible NOD2 interacts with 3-O-sulfogalactoceramide and regulates vesicular homeostasis. In: Biochimica et Biophysica Acta - General Subjects. 2013 ; Vol. 1830, No. 11. pp. 5277-5286.
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