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J Biochem. 2010 Jul;148(1):93-101. doi: 10.1093/jb/mvq036. Epub 2010 Apr 7.

Dibutyryl-cAMP up-regulates nur77 expression via histone modification during neurite outgrowth in PC12 cells.

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  • 1Laboratory of Neurobiology, Department of Life Science and Biotechnology, Faculty of Chemistry, Materials, and Bioengineering and Strategic Research Base, Kansai University, 3-3-35, Yamate-cho, Suita, Osaka 564-8680, Japan.

Abstract

An elevated level of cyclic AMP (cAMP) within cells activates gene expression through the cAMP-PKA-CREB pathway. Among the CREB target genes, some immediate early genes exist that are responsive to cAMP including the nur77 and c-fos genes. Treatment with dibutyryl-cAMP (dbcAMP) as well as nerve growth factor (NGF) induces neurite outgrowth in PC12 cells. Here, we report that acetylation of histone H3 was gradually stimulated after treatment with dbcAMP in PC12 cells and peaked 1 h after treatment. As the result of reverse transcription-polymerase chain reaction (RT-PCR) and quantitative real-time PCR (qPCR) experiments, both nur77 and c-fos gene expression were found to have peak 1 h after treatment. Knock-down with siRNA against nur77 mRNA inhibited the neurite outgrowth induced by dbcAMP, whereas knock-down with siRNA against c-fos mRNA did not inhibit the dbcAMP-induced neurite outgrowth. A chromatin immunoprecipitation (ChIP) assay revealed that the nur77 gene was associated with the acetylated Lys14 of histone H3 after treatment with dbcAMP. However, the amount of c-fos gene associated with acetylated histone H3 was not changed after treatment with dbcAMP. These results suggest that the expression of nur77, which is essential for the neuronal differentiation induced by dbcAMP, is up-regulated via dbcAMP-induced acetylation of the Lys14 of histone H3 in PC12 cells.

PMID:
20375114
[PubMed - indexed for MEDLINE]
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