Eukaryotic translation initiation factor 6 is a novel regulator of reactive oxygen species-dependent megakaryocyte maturation

J Thromb Haemost. 2015 Nov;13(11):2108-18. doi: 10.1111/jth.13150. Epub 2015 Oct 23.

Abstract

Background: Ribosomopathies constitute a class of inherited disorders characterized by defects in ribosome biogenesis and function. Classically, bone marrow (BM) failure is a clinical symptom shared between these syndromes, including Shwachman-Bodian-Diamond syndrome (SBDS). Eukaryotic translation initiation factor 6 (eIF6) is a critical translation factor that rescues the quasilethal effect of the loss of the SBDS protein.

Objectives: To determine whether eIF6 activity is necessary for BM development.

Methods: We used eIF6(+/-) mice and primary BM megakaryocytes to investigate the involvement of eIF6 in the regulation of hematopoiesis.

Results: We provide evidence that reduced eIF6 expression negatively impacts on megakaryopoiesis. We show that inhibition of eIF6 leads to a reduction in cell size and mean ploidy level of megakaryocytes and a delay in megakaryocyte maturation by blocking the G1 /S transition. Consistent with this phenotype, only few megakaryocyte-forming proplatelets were found in eIF6(+/-) cells. We also discovered that, in eIF6(+/-) cells, the steady-state abundance of mitochondrial respiratory chain complex I-encoding mRNAs is decreased, resulting in decreased reactive oxygen species (ROS) production. Intriguingly, connectivity map analysis showed that eIF6-mediated changes overlap with specific translational inhibitors. eIF6 is a translation factor acting downstream of insulin/phorbol 12-myristate 13-acetate (PMA) stimulation. PMA treatment significantly restored eIF6(+/-) megakaryocyte maturation, indicating that activation of eIF6 is essential for the rescue of the phenotype.

Conclusions: Taken together, our results show a role for eIF6-driven translation in megakaryocyte development, and unveil the novel connection between translational control and ROS production in this cell subset.

Keywords: initiation factors; megakaryocytes; platelets; reactive oxygen species; thrombocytopenia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / metabolism
  • Bone Marrow Diseases / metabolism
  • Cell Size
  • Cells, Cultured
  • Chromatin Assembly and Disassembly / physiology
  • Down-Regulation
  • Electron Transport Complex I / biosynthesis
  • Electron Transport Complex I / genetics
  • Exocrine Pancreatic Insufficiency / metabolism
  • G1 Phase / physiology
  • Lipomatosis / metabolism
  • Male
  • Megakaryocytes / metabolism
  • Megakaryocytes / pathology
  • Mice
  • Mice, Inbred C57BL
  • Peptide Initiation Factors / deficiency
  • Peptide Initiation Factors / genetics
  • Peptide Initiation Factors / physiology*
  • Phenotype
  • Ploidies
  • Protein Biosynthesis / physiology
  • RNA, Messenger / biosynthesis
  • Reactive Oxygen Species / metabolism*
  • Ribosome Subunits, Large, Eukaryotic / metabolism
  • Shwachman-Diamond Syndrome
  • Tetradecanoylphorbol Acetate / pharmacology
  • Thrombopoiesis / physiology*

Substances

  • Itgb4bp protein, mouse
  • Peptide Initiation Factors
  • RNA, Messenger
  • Reactive Oxygen Species
  • Electron Transport Complex I
  • Tetradecanoylphorbol Acetate