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Control of nuclear-cytoplasmic shuttling of Ankrd54 by PKCδ

Amy L Samuels1, Alison Louw1, Reza Zareie2

  • 1Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, the University of Western Australia, Nedlands, WA 6009, Australia.

World Journal of Biological Chemistry
|September 20, 2017
PubMed
Abstract

Insights

Phosphorylation by PKCδ regulates Ankrd54

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Ankrd54 is a protein involved in nuclear-cytoplasmic transport.
  • Phosphorylation is a key post-translational modification regulating protein function and localization.

Purpose of the Study:

  • To investigate the role of phosphorylation in Ankrd54's subcellular localization.
  • To identify kinases that regulate Ankrd54 phosphorylation and localization.

Main Methods:

  • HEK293T cells were treated with kinase activators (PMA, calyculin A, staurosporin).
  • Ankrd54 was expressed with or without Lyn tyrosine kinase variants.
  • Subcellular localization was assessed via immunofluorescence and subcellular fractionation.
  • Phosphorylation was analyzed using Phos-tag gel assays and MRM proteomics.

Main Results:

  • Phorbol 12-myristate 13-acetate (PMA) treatment induced Ankrd54 nuclear export and increased its phosphorylation.
  • PKCδ activation specifically promoted Ankrd54 phosphorylation and cytoplasmic localization.
  • Mutations of serine residues (Ser14, 17, 18, 19) reduced PMA-induced effects.
  • Phosphorylation at Ser18 was confirmed by MRM analysis.
  • PMA stimulation enhanced Ankrd54 interaction and co-localization with Lyn tyrosine kinase.

Conclusions:

  • PKCδ-mediated phosphorylation is a key regulator of Ankrd54's nuclear-cytoplasmic shuttling.
  • Phosphorylation influences Ankrd54's interaction with Lyn tyrosine kinase.
Keywords:
Ankrd54BtkLynPKCδPhosphorylation

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