Functional whole-genome analysis identifies Polo-like kinase 2 and poliovirus receptor as essential for neuronal differentiation upstream of the negative regulator alphaB-crystallin

J Biol Chem. 2009 Nov 13;284(46):32053-65. doi: 10.1074/jbc.M109.009324. Epub 2009 Aug 21.

Abstract

This study aimed at identifying transcriptional changes associated to neuronal differentiation induced by six distinct stimuli using whole-genome microarray hybridization analysis. Bioinformatics analyses revealed the clustering of these six stimuli into two categories, suggesting separate gene/pathway dependence. Treatment with specific inhibitors demonstrated the requirement of both Janus kinase and microtubule-associated protein kinase activation to trigger differentiation with nerve growth factor (NGF) and dibutyryl cAMP. Conversely, activation of protein kinase A, phosphatidylinositol-3-kinase alpha, and mammalian target of rapamycin, although required for dibutyryl cAMP-induced differentiation, exerted a negative feedback on NGF-induced differentiation. We identified Polo-like kinase 2 (Plk2) and poliovirus receptor (PVR) as indispensable for NGF-driven neuronal differentiation and alphaB-crystallin (Cryab) as an inhibitor of this process. Silencing of Plk2 or PVR blocked NGF-triggered differentiation and Cryab down-regulation, while silencing of Cryab enhanced NGF-induced differentiation. Our results position both Plk2 and PVR upstream of the negative regulator Cryab in the pathway(s) leading to neuronal differentiation triggered by NGF.

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Differentiation
  • Cells, Cultured
  • Computational Biology
  • Gene Expression Profiling
  • Genome*
  • Humans
  • Mice
  • Nerve Growth Factor / pharmacology
  • Neurons / cytology*
  • Neurons / drug effects
  • Neurons / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Receptors, Virus / genetics
  • Receptors, Virus / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • alpha-Crystallin B Chain / genetics
  • alpha-Crystallin B Chain / metabolism*

Substances

  • RNA, Messenger
  • Receptors, Virus
  • alpha-Crystallin B Chain
  • poliovirus receptor
  • Nerve Growth Factor
  • Protein Serine-Threonine Kinases
  • Plk2 protein, rat