The RNA-binding protein HuD is required for GAP-43 mRNA stability, GAP-43 gene expression, and PKC-dependent neurite outgrowth in PC12 cells

C. D. Mobarak, K. D. Anderson, M. Morin, A. Beckel-Mitchener, S. L. Rogers, H. Furneaux, P. King, N. I. Perrone-Bizzozero

Research output: Contribution to journalArticle

106 Scopus citations

Abstract

The RNA-binding protein HuD binds to a regulatory element in the 3′ untranslated region (3 ′ UTR) of the GAP-43 mRNA. To investigate the functional significance of this interaction, we generated PC12 cell lines in which HuD levels were controlled by transfection with either antisense (pDuH) or sense (pcHuD) constructs, pDuH-transfected cells contained reduced amounts of GAP-43 protein and mRNA, and these levels remained low even after nerve growth factor (NGF) stimulation, a treatment that is normally associated with protein kinase C (PKC)-dependent stabilization of the GAP-43 mRNA and neuronal differentiation. Analysis of GAP-43 mRNA stability demonstrated that the mRNA had a shorter half-life in these cells. In agreement with their deficient GAP-43 expression, pDuH cells failed to grow neurites in the presence of NGF or phorbol esters. These cells, however, exhibited normal neurite outgrowth when exposed to dibutyryl-cAMP, an agent that induces outgrowth independently from GAP-43. We observed opposite effects in pcHuD-transfected cells. The GAP-43 mRNA was stabilized in these cells, leading to an increase in the levels of the GAP-43 mRNA and protein. pcHuD cells were also found to grow short spontaneous neurites, a process that required the presence of GAP-43. In conclusion, our results suggest that HuD plays a critical role in PKC-mediated neurite outgrowth in PC12 cells and that this protein does so primarily by promoting the stabilization of the GAP-43 mRNA.

Original languageEnglish (US)
Pages (from-to)3191-3203
Number of pages13
JournalMolecular biology of the cell
Volume11
Issue number9
DOIs
StatePublished - 2000

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Mobarak, C. D., Anderson, K. D., Morin, M., Beckel-Mitchener, A., Rogers, S. L., Furneaux, H., King, P., & Perrone-Bizzozero, N. I. (2000). The RNA-binding protein HuD is required for GAP-43 mRNA stability, GAP-43 gene expression, and PKC-dependent neurite outgrowth in PC12 cells. Molecular biology of the cell, 11(9), 3191-3203. https://doi.org/10.1091/mbc.11.9.3191