Hypothyroidism-associated missense mutation impairs NADPH oxidase activity and intracellular trafficking of Duox2

Ágnes Donkó, Stanislas Morand, Agnieszka Korzeniowska, Howard E. Boudreau, Melinda Zana, L. Hunyady, M. Geiszt, Thomas L. Leto

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

In the thyroid gland Duox2-derived H2O2 is essential for thyroid hormone biosynthesis. Several patients were identified with partial or severe iodide organification defects caused by mutation in the gene for Duox2 or its maturation factor, DuoxA2. A Duox2-deficient (Duox2thyd) mouse model enabled in vivo investigation of its critical function in thyroid tissues, but its roles proposed in host defense or other innate responses in nonthyroid tissues remain less certain. These mice carry a spontaneous DUOX2 missense mutation, a T→G transversion, in exon 16 that changes the highly conserved valine 674 to glycine and results in severe congenital hypothyroidism. The exact mechanism underlying the effects of the V674G mutation has not been elucidated at the molecular or cellular level. To determine how the V674G mutation leads to congenital hypothyroidism, we introduced the same mutation into human Duox2 or Duox1 cDNAs and expressed them in HEK-293 cells stably expressing the corresponding DuoxA proteins. We found that the valine→glycine mutant Duox proteins fail to produce H2O 2, lose their plasma membrane localization pattern, and are retained within the endoplasmic reticulum. The Duox2 mutant binds to DuoxA2, but appears to be unstable owing to this retention. Immunohistochemical staining of Duox2 in murine salivary gland ducts showed that Duox2 in mutant mice loses its condensed apical plasma membrane localization pattern characteristic of wild-type Duox2 and accumulates in punctate vesicular structures within cells. Our findings demonstrate that changing the highly conserved valine 674 in Duox2 leads to impaired subcellular targeting and reactive oxygen species release required for hormonogenesis, resulting in congenital hypothyroidism.

Original languageEnglish
Pages (from-to)190-200
Number of pages11
JournalFree Radical Biology and Medicine
Volume73
DOIs
Publication statusPublished - 2014

Fingerprint

NADPH Oxidase
Valine
Missense Mutation
Cell membranes
Hypothyroidism
Congenital Hypothyroidism
Tissue
Mutation
Biosynthesis
Iodides
Thyroid Hormones
Ducts
Glycine
Exons
Reactive Oxygen Species
Thyroid Gland
Proteins
Complementary DNA
Genes
Cell Membrane

Keywords

  • Duox
  • Free radicals
  • Hypothyroidism
  • Mutation
  • NADPH oxidase

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Hypothyroidism-associated missense mutation impairs NADPH oxidase activity and intracellular trafficking of Duox2. / Donkó, Ágnes; Morand, Stanislas; Korzeniowska, Agnieszka; Boudreau, Howard E.; Zana, Melinda; Hunyady, L.; Geiszt, M.; Leto, Thomas L.

In: Free Radical Biology and Medicine, Vol. 73, 2014, p. 190-200.

Research output: Contribution to journalArticle

Donkó, Ágnes ; Morand, Stanislas ; Korzeniowska, Agnieszka ; Boudreau, Howard E. ; Zana, Melinda ; Hunyady, L. ; Geiszt, M. ; Leto, Thomas L. / Hypothyroidism-associated missense mutation impairs NADPH oxidase activity and intracellular trafficking of Duox2. In: Free Radical Biology and Medicine. 2014 ; Vol. 73. pp. 190-200.
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