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

Research output: Contribution to journalArticle

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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
Pages (from-to)3191-3203
Number of pages13
JournalMolecular Biology of the Cell
Volume11
Issue number9
StatePublished - Dec 1 2000
Externally publishedYes

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GAP-43 Protein
RNA-Binding Proteins
PC12 Cells
RNA Stability
Protein Kinase C
Gene Expression
Messenger RNA
Proteins
Nerve Growth Factor
Neurites
Neuronal Outgrowth
3' Untranslated Regions
Phorbol Esters
Transfection
Half-Life

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Mobarak, C. D., Anderson, K. D., Morin, M., Beckel-Mitchener, A., Rogers, S. L., Furneaux, H., ... 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.

The RNA-binding protein HuD is required for GAP-43 mRNA stability, GAP-43 gene expression, and PKC-dependent neurite outgrowth in PC12 cells. / Mobarak, C. D.; Anderson, Kimberly D; Morin, M.; Beckel-Mitchener, A.; Rogers, S. L.; Furneaux, H.; King, P.; Perrone-Bizzozero, N. I.

In: Molecular Biology of the Cell, Vol. 11, No. 9, 01.12.2000, p. 3191-3203.

Research output: Contribution to journalArticle

Mobarak, CD, Anderson, KD, Morin, M, Beckel-Mitchener, A, Rogers, SL, Furneaux, H, King, P & Perrone-Bizzozero, NI 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, vol. 11, no. 9, pp. 3191-3203.
Mobarak CD, Anderson KD, Morin M, Beckel-Mitchener A, Rogers SL, Furneaux H et al. 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. 2000 Dec 1;11(9):3191-3203.
Mobarak, C. D. ; Anderson, Kimberly D ; Morin, M. ; Beckel-Mitchener, A. ; Rogers, S. L. ; Furneaux, H. ; King, P. ; Perrone-Bizzozero, N. I. / The RNA-binding protein HuD is required for GAP-43 mRNA stability, GAP-43 gene expression, and PKC-dependent neurite outgrowth in PC12 cells. In: Molecular Biology of the Cell. 2000 ; Vol. 11, No. 9. pp. 3191-3203.
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