Abstract
Neuronal outgrowth is guided by both extrinsic and intrinsic factors, involving transcriptional regulation. The acetylation of histones and transcription factors, which facilitates promoter accessibility, ultimately promotes transcription, and depends on the balance between histone deacetylases (HDACs) and histone acetyltransferases (HATs) activities. However, a critical function for specific acetylation modifying enzymes in neuronal outgrowth has yet to be investigated. To address this issue, we have used an epigenetic approach to facilitate gene expression in neurons, by using specific HDAC inhibitors. Neurons treated with a combination of HDAC and transcription inhibitors display an acetylation and transcription-dependent increase in outgrowth and a reduction in growth cone collapse on both ‘permissive’ (poly-D-lysine, PDL) and ‘non-permissive’ substrates (myelin and chondroitin sulphate proteoglycans (CSPGs)). Next, we specifically show that the expression of the histone acetyltransferases CBP/p300 and P/CAF is repressed in neurons by inhibitory substrates, whereas it is triggered by HDAC inhibition on both permissive and inhibitory conditions. Gene silencing and gain of function experiments show that CBP/p300 and P/CAF are key players in neuronal outgrowth, acetylate histone H3 at K9-14 and the transcription factor p53, thereby initiating a pro-neuronal outgrowth transcriptional program. These findings contribute to the growing understanding of transcriptional regulation in neuronal outgrowth and may lay the molecular groundwork for the promotion of axonal regeneration after injury.
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Abbreviations
- CGN:
-
cerebellum granular neuron
- CBP:
-
CREB-binding protein
- CSPG:
-
chondroitin sulphate proteoglycan
- GAP-43:
-
growth-associated protein-43
- HAT:
-
histone acetyltransferase
- HDAC:
-
histone deacetylase
- PB:
-
sodium phenyl butyrate
- P/CAF:
-
p300/CBP-associated factor
- PDL:
-
poly-D-lysine
- RCN:
-
rat cortical neuron
- SAHA:
-
suberoylanilide hydroxamic acid
- TSA:
-
trichostatin A
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Acknowledgements
We thank Dr. Maria Laura Avantaggiati for providing the p53 acetylation mutant plasmid DNAs, and Dr. Katsuhide Miyake for providing the CBP/p300 RNAi. We also thank Dr. Richard Goodman for providing the CBP plasmid, Dr. Yoshihiro Nakatani for providing the P/CAF plasmid and Dr. William Sellers for providing the p300 plasmid. This work was supported by the Hertie Foundation; the Fortune Grant, University of Tuebingen, the NIH R21 NS052640 and the DFG DI 1497/1-1 grants (all granted to Simone Di Giovanni).
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Gaub, P., Tedeschi, A., Puttagunta, R. et al. HDAC inhibition promotes neuronal outgrowth and counteracts growth cone collapse through CBP/p300 and P/CAF-dependent p53 acetylation. Cell Death Differ 17, 1392–1408 (2010). https://doi.org/10.1038/cdd.2009.216
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DOI: https://doi.org/10.1038/cdd.2009.216
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