Identification of Novel MAGE-G1-Interacting Partners in Retinoic Acid-Induced P19 Neuronal Differentiation Using SILAC-Based Proteomics

Sci Rep. 2017 Apr 4;7:44699. doi: 10.1038/srep44699.

Abstract

MAGE-G1 is a protein plays role in the early process of neurogenesis. However, the fundamental roles MAGE-G1 played in neurogenesis have not yet been completely understood. Finding the partners MAGE-G1 interacting with will surely contribute to the function study of MAGE-G1. In this study, using Stable Isotope Labeling by Amino acids in Cell culture-immunoprecipitation quantitative proteomics, we screened the interacting proteins of MAGE-G1 during retinoic acid -induced neuronal differentiation of P19 cells and firstly found that FSCN1 and VIME were potential novel MAGE-G1-interacting proteins. Then, the interaction between overexpressed MAGE-G1 and FSCN1 or VIME was validated by GST-pull down assay in bacteria and by co-immunoprecipitation assay in COS7 cells. Endogenous co-immunoprecipitation assay further confirmed that MAGE-G1 interacted with FSCN1 or VIME in P19 cells after a 6-day retinoic acid-induced neuronal differentiation. Those results provide a functional linkage between MAGE-G1 and FSCN1 or VIME and may facilitate a better understanding of the fundamental aspects of MAGE-G1 during neurogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Differentiation / drug effects
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Gene Expression Regulation, Neoplastic*
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Isotope Labeling
  • Melanoma-Specific Antigens / genetics*
  • Melanoma-Specific Antigens / metabolism
  • Mice
  • Microfilament Proteins / genetics*
  • Microfilament Proteins / metabolism
  • Neoplastic Stem Cells / cytology
  • Neoplastic Stem Cells / drug effects
  • Neoplastic Stem Cells / metabolism
  • Neurogenesis / genetics
  • Neurons / cytology
  • Neurons / drug effects*
  • Neurons / metabolism
  • Protein Binding
  • Proteomics / methods
  • Receptors, Odorant / genetics*
  • Receptors, Odorant / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction
  • Tretinoin / pharmacology*
  • Vimentin / genetics*
  • Vimentin / metabolism

Substances

  • Melanoma-Specific Antigens
  • Microfilament Proteins
  • Receptors, Odorant
  • Recombinant Fusion Proteins
  • Vim protein, mouse
  • Vimentin
  • fascin1 protein, mouse
  • Tretinoin
  • Glutathione Transferase