Impaired activity of CCA-adding enzyme TRNT1 impacts OXPHOS complexes and cellular respiration in SIFD patient-derived fibroblasts

Urszula Liwak-Muir; Hapsatou Mamady; Turaya Naas; Quinlan Wylie; Skye McBride; Matthew Lines; Jean Michaud; Stephen D Baird; Pranesh K Chakraborty; Martin Holcik

doi:10.1186/s13023-016-0466-3

Impaired activity of CCA-adding enzyme TRNT1 impacts OXPHOS complexes and cellular respiration in SIFD patient-derived fibroblasts

Orphanet J Rare Dis. 2016 Jun 18;11(1):79. doi: 10.1186/s13023-016-0466-3.

Authors

Urszula Liwak-Muir¹, Hapsatou Mamady¹, Turaya Naas^{1

2}, Quinlan Wylie¹, Skye McBride¹, Matthew Lines^{1

3}, Jean Michaud⁴, Stephen D Baird¹, Pranesh K Chakraborty^{5

6}, Martin Holcik^{1

3}

Affiliations

¹ Molecular Biomedicine Program, Children's Hospital of Eastern Ontario Research Institute, 401 Smyth Road, Ottawa, ON, K1H 8L1, Canada.
² Newborn Screening Ontario, Children's Hospital of Eastern Ontario, Ottawa, ON, Canada.
³ Department of Pediatrics, University of Ottawa, Ottawa, Ontario, K1H 8M5, Canada.
⁴ Department of Pathology and Laboratory Medicine, Children's Hospital of Eastern Ontario and University of Ottawa, Ottawa, Ontario, Canada.
⁵ Department of Pediatrics, University of Ottawa, Ottawa, Ontario, K1H 8M5, Canada. pchakraborty@cheo.on.ca.
⁶ Newborn Screening Ontario, Children's Hospital of Eastern Ontario, Ottawa, ON, Canada. pchakraborty@cheo.on.ca.

Abstract

Background: SIFD (Sideroblastic anemia with B-cell immunodeficiency, periodic fevers, and developmental delay) is a novel form of congenital sideroblastic anemia associated with B-cell immunodeficiency, periodic fevers, and developmental delay caused by mutations in the CCA-adding enzyme TRNT1, but the precise molecular pathophysiology is not known.

Results: We show that the disease causing mutations in patient-derived fibroblasts do not affect subcellular localization of TRNT1 and show no gross morphological differences when compared to control cells. Analysis of cellular respiration and oxidative phosphorylation (OXPHOS) complexes demonstrates that both basal and maximal respiration rates are decreased in patient cells, which may be attributed to an observed decrease in the abundance of select proteins of the OXPHOS complexes.

Conclusions: Our data provides further insight into cellular pathophysiology of SIFD.

Keywords: Mitochondria; OXPHOS; SIFD; TRNT1; Translation.

MeSH terms

Anemia, Sideroblastic / genetics
Anemia, Sideroblastic / metabolism*
Blotting, Western
Cell Respiration / genetics
Cell Respiration / physiology*
Cells, Cultured
Female
Fibroblasts / metabolism*
Fluorescent Antibody Technique
Humans
Membrane Potential, Mitochondrial
Microscopy, Electron, Transmission
Mitochondria / metabolism
Mutation
Nucleotidyltransferases / genetics
Nucleotidyltransferases / metabolism*
Oxidative Phosphorylation

Substances

Nucleotidyltransferases
TRNT1 protein, human